Startseite Lebenswissenschaften Value-added analysis of Lactobacillus acidophilus cell encapsulation using Eucheuma cottonii by freeze-drying and spray-drying
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Value-added analysis of Lactobacillus acidophilus cell encapsulation using Eucheuma cottonii by freeze-drying and spray-drying

  • Silvia Oktavia Nur Yudiastuti EMAIL logo , Roni Kastaman , Een Sukarminah und Efri Mardawati
Veröffentlicht/Copyright: 27. April 2022
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Open Agriculture
Aus der Zeitschrift Open Agriculture Band 7 Heft 1

Abstract

The added value of Lactobacillus acidophilus encapsulation due to its production process is one of the first things that need to be known in calculating techno-economic analysis to determine the sustainability of its downstream. The result of value-added analysis plays an important role in determining which formulation, encapsulate material, and production process should be used. The inoculum media used were skimmed milk and whole milk, the coating material used in this study was Eucheuma cottonii, while the process variations used were spray-drying and freeze-drying methods. There were four treatments in the research that analyzed their added value. Determination of the best treatment was carried out through the scoring method on the value of cell viability, cell morphology, encapsulation effectiveness, process costs, cost of supporting materials, and added value of the product. Added value analysis was carried out using the Hayami and Kawagoe methods. Based on the research results, it can be concluded that added value is most influenced by process costs. The treatments selected were E. cottonii coating material, fresh milk inoculation medium, and spray-drying encapsulation method with an overall score of 22.

Keywords: formulation; Hayami and Kawagoe; hydrated cell; process cost; techno-economic analysis

1 Introduction

Probiotics are live cells that are consumed as part of the daily diet and can provide healthful effects through their metabolic processes in the human colon [1]. The healthful effect of probiotics occurs through the lactose metabolism process by probiotic cells in the digestive system into organic acids such as butyric acid, lactic acid, and acetic acid [2,3]. These organic acids can help bowel activity by stimulating peristalsis and stimulating the immune system to have resistance to infection and cancer [4]. Probiotics can provide health effects if their content in the food consumed is at least 107 CFU/g and a minimum of 105 CFU/g in the digestive tract system [5,6,7]. Lactobacillus is the most widely developed probiotic to be produced among other probiotic strains because it is homofermentative, and is present naturally in the human colon [8]. Lactobacillus acidophilus is the most acid resistant among other species in the Lactobacillus genus [9]. Several studies have focused on the characterization of new strains, formulations, ready-to-consume probiotic dosage forms, and their stability during storage and in the human digestive tract. The results showed that probiotics were more stably produced in dry conditions [10,11,12,13].

L. acidophilus encapsulation result is expected to be applied in small and medium enterprises so that the raw materials ease consideration, raw materials costs, production costs, high productivity, production ease, and added value of products are things that need attention. The L. acidophilus encapsulation through drying is referred to as the physicomechanical method which has the advantage that it does not involve polymerization reaction considering that both the encapsulation material and the inoculation media are polymers so that in general only shape formation occurs [14]. Spray-drying is a dry food preparation production method that is widely used in the food industry because it has high effectiveness and productivity even though it has disadvantages due to the high use of heat [15].

Several studies of spray-dried encapsulation method have been carried out on L. acidophilus cells using inulin [12,16], acacia gum [17], locust bean gum [18], starch [19], soy protein [17], whey protein [16,20,21,22], skim milk [23], chitosan [22,24], and alginate [22]. These studies show that the advantages of spray-dried encapsulation method among others mass production are continuous process, material monodispersity, well established in the food industry for other applications, and inexpensive; however, this method still has drawbacks, among them is the loss of cell viability due to high temperature and mostly used an aqueous suspension [25].

The use of high temperatures can cause osmotic stress on L. acidophilus [11]. The drawbacks of spray-drying can be overcome, one of which is through the use of an L. acidophilus coating agent with a prebiotic called co-encapsulation [26]. Seaweed is an abundant source of prebiotics that can be used as a coating for probiotics because it contains galactooligosaccharides [27]. Seaweed is easy to obtain considering that the amount of oceans on earth reaches 71%. Eucheuma cottonii has a kappa-carrageenan gel type so it has a stronger gel strength than other genera [28]. In addition, Indonesia is the largest E. cottonii producer in the world; Indonesia’s ocean area is 70% of Indonesia’s total area so business activists will not have trouble finding E. cottonii.

An alternative drying method used for temperature-sensitive food products is freeze-drying [29]. Several studies on freeze-dried encapsulation method have been carried out on L. acidophilus cells using starch, soy protein [30,31], casein [31], whey protein [31], and skim milk [31]. Based on these studies, the advantage of the freeze-dried encapsulation method was the excellent final hydrated material suitable for most food applications and suitable for sensitive materials probiotics [25]. Meantime, the disadvantages were high cost, cell damage with eventual crystal formation if the method is not carried out correctly, and the eventual need for cryoprotectants [25]. The longer time is a drawback of the freeze-drying method because it requires more energy which in turn increases the production costs [32]. One of the ways to reduce production costs in the freeze-drying method is by replacing the skim milk inoculum with fresh milk, which is easier for the public or business actors to find.

The main objective of this research was to optimize the hydrated L. acidophilus value-added in dry powder preparation by comparing two types of production methods, one type of coating material, and two types of inoculum media. This was done as an experimental data simulation for industrial scale-up preparation. Simulation of L. acidophilus probiotic production was carried out from cell rejuvenation, inoculation with skim or whole milk, coating with E. cottonii, drying by spray-drying or freeze-drying, analysis of production effectiveness, cell viability, cell morphology, and value-added analysis. The determination of rating scale for the selected treatment was carried out at the end of the research.

2 Material and methods

2.1 Experimental site and material description

The experiment was carried out from 2018 to 2019 at the Department of Food Technology Padjadjaran University, Indonesia. The probiotic used was L. acidophilus ATCC 4356 which was purchased from the Laboratory of Microbiology, Department of Biology, Padjadjaran University. All the analytical chemicals were purchased from Sigma-Aldrich (USA), Merck (Germany), and Himedia laboratories Pvt Ltd. Fresh milk was purchased from the Faculty of Husbandry, Universitas Padjadjaran, while skim milk was purchased from NZMP, New Zeland. All reagents were analytical grade, deionized water was used for sample preparation and analyses.

2.2 Treatment, experimental design, and statistical analysis

Treatment includes two levels of drying method (freeze-drying and spray-drying), two levels of inoculum media (skim milk and pasteurized fresh milk), and one level of coating material (E. cottonii). The combination of treatment was FDS (freeze-drying – skim milk – E. cottonii), FDF (freeze-drying – pasteurized fresh milk – E. cottonii), SDS (spray-drying – skim milk – E. cottonii), and SDF (spray-drying – pasteurized fresh milk – E. cottonii). As value-added control for hydrated L. acidophilus, the value-added calculation was done to pure culture as raw materials of hydrated L. acidophilus. The composition used was 10% cell culture, 20% encapsulated material, and 10% inoculum media. The experiment was laid out in Randomized Block Design (RBD), replicated three times. All data obtained were subjected to analysis of variance (ANOVA) using the statistical Microsoft excel. Significant means were separated using Duncan’s multiple comparison test at a 5% level of probability.

2.3 L. acidophilus preparation

The pure culture of Lactobacillus acidophilus was rejuvenated and propagated in a test tube with a sloping method using MRS agar media. Lactobacillus acidophilus was then incubated at 38°C for 48 h. Rejuvenation cultures were then decomposed using 0.5–2 mL of physiological NaCl solution in each test tube. The turbidity was checked according to McFarland 3 at λ = 600 nm and absorbance ±0.616 which is equivalent to the number of colonies (3.0 × 108 CFU/mL) on a spectrophotometer. The culture solution of as much as 10% was inoculated into pasteurized fresh milk as inoculum media. The inoculum was incubated for 4 h at 40°C [26].

2.4 L. acidophilus encapsulated with freeze-dried and spray-dried method

The culture was suspended with E. cottonii fluid mixture as coating material as much 20% part from inoculum media. E. cottonii fluid mixture was prepared by pulverizing one part of E. cottonii with one part of deionized water. The suspended mixture was homogenized with a magnetic stirrer, then placed in a sterile glass beaker and covered with plastic wrap. The feed liquid composition as the suspended mixture was 1/13 part of culture solution, 10/13 part of inoculum media, and 2/13 part of coating material. The suspended mixture was filled in a freeze-drying chamber, wrapped with aluminum foil, and stored overnight in a freezing cabinet at −60°C. Thereafter, the freeze suspended mixture in the chamber was freeze-dried for 24–48 h, at −55°C and 0.050 mBar vacuum. Meanwhile, for the spray-dried method, the suspended culture mixture was hydrated by inlet and outlet temperature at 100 and 75°C with a flow rate at 20 g/min. The dry powder was collected in a sterile tank attached to the bottom of the cyclone. After freeze-drying and spray-drying, the hydrated L. acidophilus was collected and stored at 4°C for further experiments [26].

2.5 Field Emission-Scanning Electron Microscope (FE-SEM) image of encapsulated L. acidophilus

The particle morphology of encapsulated L. acidophilus was examined using a SEM (SEM S-4800, Hitachi, Japan). The accelerating voltage was 10 kV. The sample was dissolved in a copper container coated with carbon. The copper container was then put into a freeze-dryer. The hydrated samples were then observed using SEM [33].

2.6 Encapsulation effectivity (EE)

The calculations were carried out by comparing results of L. acidophilus suspended weight and weight of hydrated L. acidophilus [34]. The encapsulation effectivity (EE) of encapsulation was calculated for L. acidophilus by

(1) EE ( % ) = ( Hydrated weight / suspended weight ) × 100 % .

2.7 Calculation of total bacteria L. acidophilus and cell viability test

1 mL of sample was diluted with 9 mL of 0.85% NaCl physiological solution, up to 10−9 aseptically. Each tube in the last three dilutions was pinched every 1 mL into a sterile petri dish. MRS medium was poured and 0.5% glacial acetic acid of 12–15 mL was added into the petri dish, and then incubated at 37°C for 48 h. The number of colonies grown on agar was counted by BAM methods [34]. The cell viability was counted using the following formulation

(2) Cell viability ( % ) = ( Count of hydrated L . a c i d o p h i l u s / Count of suspended L . a c i d o p h i l u s suspended weight ) × 100 % .

2.8 Value-added analysis

The analysis was conducted according to Hayami and Kawagoe method [35] with an example calculation as presented by Yudiastuti et al. [36,37].

2.9 The determination of scores criteria at observed parameters in searching the best treatment

The determined score was based on the upper and lower limits of the value obtained for each observed parameter. This was done to provide a specific score based on the quality value of each parameter tested on the cell encapsulation results. The scores used in this research to determine the best treatment are presented in Table 1.

Table 1

The scoring value criteria of parameters observed

Observation recapitulation Scoring value
4 3 2 1
L. acidophilus (CFU/g) >12.01 11.51–12.00 11.01–11.50 10.51–11.00
Viability (%) >94.0 93.2–92 91.9–90.0 <89.9
EE (%) >10 10.1–8 7.9–5 <4.9
Added value (IDR/g) >13,000 10,001–13,000 5,001–10,000 <5,000
Process cost <50 50.1–100 100.1–150 >150.1
Total raw material cost <300 301–459.9 450–499.9 >500

2.10 Physicochemical characterization

Characterization was carried out on the product with the highest score based on scoring data results. Characterizations that were carried out were moisture content, water activity (aw), hygroscopicity, dissolution, and human gastrointestinal invitro test.

2.10.1 Moisture content and water activity [38]

Moisture content was analyzed using the gravimetric method. A total of 2 g sample was placed in weighted solidified aluminum pans. The sample and pan were put into the oven at 105°C for 5 h and then weighed until a constant weight was obtained. Moisture content was calculated according to the formula [38]:

(3) % Moisture content = [ ( Initial weight of sample − Final weight of sample ) / Initial weight of sample ] × 100 .

Water activity was measured using Aqua Lab Water Activity meter (Aqua Lab, USA)

2.10.2 Hygroscopicity [39]

A total of 1 g sample was placed into weighted solidified aluminum pans. The sample and pan were placed into a desiccator equilibrated at 75% relative humidity containing saturated sodium chloride. The sample was kept for 7 days and hygroscopicity was calculated using the following formula:

(4) Hygroscopicity ( gH 2 O / g sample ) = [ ( Moisture content of sample after storage − Initial moisture content of sample ) / Initial moisture content of sample ] × 100 .

2.10.3 Dissolution [39]

The dissolution capacity test was done by dissolved 1 g sample into 50 mL of distilled water. The sample solution was then stirred using a magnetic stirrer of 2 mm × 7 mm stirring bar at medium speed. The time required by encapsulation powder to complete dissolution was recorded in minutes.

2.10.4 In vitro test of human gastrointestine [7]

A total of 10 g sample was dissolved in 90 mL of NaCl solution (0.5% w/v) and glucose anhydrate solution (20% w/v). The sample was conditioned in a gastric juices environment for 2 h at 37°C and was stirred constantly with a magnetic stirrer. The gastric juice contained a mixture of pepsin (3 g/L), lipase (0.9 mg/L), and the pH was adjusted to a value of 2 by adding 1 N HCl. After 2 h, the solution was incubated in a pancreatic environment by adding bile salts (10 g/L) and pancreatic fluid (1 g/L) and remained incubated at 37°C for 2 h, the pH was adjusted to a value of 4.5. The pH was further adjusted to 6.5 the incubation continued for another for 2 h. The total incubation time was 6 h. The sample was tested for the total amount of L. acidophilus.

3 Result

3.1 Morphological result of hydrated L. acidophilus

SEM analysis aims to determine the microstructure of hydrated L. acidophilus. Microstructural information can provide information as a reference for descriptive observations regarding the impact of different methods production, inoculum medium type, and coating material used to the morphological cross-section. Based on the research results, images of the morphological analysis of hydrated L. acidophilus can be seen in Figure 1.

Figure 1 Hydrated L. acidophilus morphology structure. (a) FDS (freeze-drying – skim milk – E. cottonii), (b) FDF (freeze-drying – pasteurized fresh milk – E. cottonii), (c) SDS (spray-drying – skim milk – E. cottonii). (d) SDF (spray-drying – pasteurized fresh milk – E. cottonii).
Figure 1

Hydrated L. acidophilus morphology structure. (a) FDS (freeze-drying – skim milk – E. cottonii), (b) FDF (freeze-drying – pasteurized fresh milk – E. cottonii), (c) SDS (spray-drying – skim milk – E. cottonii). (d) SDF (spray-drying – pasteurized fresh milk – E. cottonii).

In the four photos, it can be seen that the size of the hydrated cell particles by spray-drying appears more uniform and smaller. This was because, in spray-drying, the particle size was determined by the micron-scale nozzle, so that the hydrated cell size particles become smaller and uniform. This is in contrast to the results of freeze-drying, where the particle size appears larger.

3.2 Process and raw materials cost of hydrated L. acidophilus production

The process production cost of hydrated L. acidophilus is presented in Table 2. The L. acidophilus hydration process was carried out by freeze-dried and spray-dried methods, the costs of the hydration process varied depending on the time it took to hydrate the L. acidophilus cells suspended. Hydration time was determined by total solid composition in L. acidophilus suspended form. Based on Table 2, the treatment with the lowest process cost was the treatment combination of spray-drying method, skim milk inoculum media, and E. cottonii encapsulation material. As a control parameter, the process cost of liquid pure culture without drying treatment, inoculum media, and coating material was calculated.

Table 2

Process cost of hydrated L. acidophilus

Process cost component Treatment
FDS FDF SDS SDF Control
Process cost in one cycle (IDR) (a) 250,000 275,022 92,000 100,000 50,000
Process time (h) (b) 60 66 0.92 1 48
Suspended material in one cycle (g) (c) 500 500 2,000 2,000 200
Unit process cost (IDR/g) (d = a/c) 500 550 46 50 250

(a) FDS (freeze-drying – skim milk – E. cottonii), (b) FDF (freeze-drying – pasteurized fresh milk – E. cottonii), (c) SDS (spray-drying – skim milk – E. cottonii). (d) SDF (spray-drying – pasteurized fresh milk – E. cottonii).

The raw materials price used was price one for the lowest price, price two for the normal price, and price three for the higher price. The raw material prices are presented in Table 3, these prices were the primary prices obtained when conducting the research. The raw material value prices were used to calculate the overall raw material price, namely cell culture, inoculum media, and coating materials required according to the formulation per unit gram of raw material. In addition, the price of pure culture raw materials per test tube used as the mother culture in the research was presented as a control raw material for calculating its added value.

Table 3

Raw material cost of hydrated L. acidophilus

Raw material component Unit price (IDR/g)
1 2 3
Skim milk 80 100 120
Distilled water 1.5 2 3.5
Fresh Milk 7 8 10
E. cottonii 32 36 40
Freeze-dried cell culture 290 461 682
Spray-dried cell culture 140 199 308
Pure culture in one test tube 250,000 350,000 450,000

3.3 Effect of treatment on various parameters observed in hydrated L. acidophilus production

Table 4 shows the treatment effect on hydrate L. acidophilus production at the total L. acidophilus cell concentrations, their viability, and the encapsulation effectiveness. Table 4 also shows the information on the L. acidophilus suspended volume, the amount of hydrate L. acidophilus, raw material costs, and processing costs. Based on Table 4, the treatment with the freeze-drying method had the highest number of cells and better viability than the treatment with the spray-drying method. However, the spray-drying method has significantly lower processing and raw material costs than the freeze-drying method. Treatment with fresh milk inoculum media resulted in a greater yield value than treatment with skim milk in both the drying methods. Table 4 also presents liquid pure culture as control (without treatment) as product value-added comparison from its raw material.

Table 4

The effect of treatment on various parameters observed

Properties Treatment
FDS FDF SDS SDF Control
L. acidophilus CFU/g) 12.2 ± 0.2bc 11.8 ± 0.03ab 11.9 ± 0.06a 11.9 ± 0.07a 9.01 ± 1.06
Viability (%) 94.2 ± 1.8%cd 91.6 ± 0.4%ab 92.5 ± 1.3%abc 92.7 ± 0.3%abc —
EE (%) 11.61 ± 2%a 11.84 ± 1%a 8.6 ± 0.7%a 10.31 ± 1%a —
Cell suspended volume (mL) 500 500 2,000 2,000 200
Final product or hydrated cell (g) 58.05 59.2 172 206.2 200
Raw material total cost (IDR/g) 494 490 281 265 4,375
Process cost (IDR/g) 500 550 46 50 250

(a) FDS (freeze-drying – skim milk – E. cottonii), (b) FDF (freeze-drying – pasteurized fresh milk – E. cottonii), (c) SDS (spray-drying – skim milk – E. cottonii). (d) SDF (spray-drying – pasteurized fresh milk – E. cottonii).

The treatments that have the same letter notation are considered the same (not significantly different) according to Duncan’s test.

3.4 Value-added calculation of hydrated L. acidophilus production

The hydrated L. acidophilus value-added calculation was carried out using Hayami and Kawagoe methods which emphasize the cost of raw materials and its process. The value-added calculation was carried out for 1 year of production because in 1 year there were 48 production cycles. Details of raw material costs, labor, working days, other contribution costs, and product prices are presented in Table 6. Other contribution costs were packaging costs, transportation costs, and production support costs. The number of workers, days of work, labor wages, and other contribution costs were made equal to see the effect of raw material costs and process costs that were influenced by product formulation and production methods. Hydrated L. acidophilus prices shown in Table 5 were the prices of similar products in the market with the highest retail prices. Table 6 shows the added value components of hydrated L. acidophilus calculated by the Hayami and Kawagoe methods. The treatment with the greatest added value was FDF (freeze-drying – pasteurized fresh milk – E. cottonii) treatment combination, but the treatment with the highest added value and profit ratio was the treatment of spray-drying method, fresh milk inoculum media, and E. cottonii encapsulation material (SDF). Control treatment has an added value of 625 IDR/g with an added value ratio of 6.25% but has a negative or no profit value.

Table 6

The effect of treatment on added value of hydrated L. acidophilus production

Parameters FDS FDF SDS SDF Control
Added value (IDR/g) 13,087 13,407 10,663 13,030 625
Added value ratio (%) 82.0 82.4 90.2 91.9 6.25
Labor share (%) 22.9 22.4 7.0 5.8 120
Profit (IDR/g) 10,087 10,407 9,913 12,280 −125
Profit level (%) 77.1 77.6 93.0 94.2 −20
Profit margin (IDR/g) 15,087 15,407 11,163 13,530 5,625
Company owner profit (%) 66.9 67.5 88.8 90.8 −2.22

(a) FDS (freeze-drying – skim milk – E. cottonii), (b) FDF (freeze-drying – pasteurized fresh milk – E. cottonii), (c) SDS (spray-drying – skim milk – E. cottonii). (d) SDF (spray-drying – pasteurized fresh milk – E. cottonii).

Table 5

Raw material price and other input contribution in production of hydrated L. acidophilus

Production in 1 year = 4 weeks × 12 months = 48 cycles
FDS FDF SDS SDF Control
Raw material (cell culture) (g) 50 50 200 200 200
Product or hydrated L. acidophilus (g) 58.05 59.2 172 206.2 200
Labor (man) 3 3 3 3 3
Days of work (in 1 week) 5 5 5 5 5
Labor wage (Man/days) 50,000 50,000 50,000 50,000 50,000
Price
Raw material
Condition 1 (IDR/g) 703 700 592 582 3,125
Condition 2 (IDR/g) 877 873 662 646 4,375
Condition 3 (IDR/g) 1,102 1,096 787 765 5,625
Other input donation (IDR) 100,000 100,000 100,000 100,000 100,000
Price of hydrated L. acidophilus or final product (IDR/g) 13,750 13,750 13,750 13,750 10,000

(a) FDS (freeze-drying – skim milk – E. cottonii), (b) FDF (freeze-drying – pasteurized fresh milk – E. cottonii), (c) SDS (spray-drying – skim milk – E. cottonii). (d) SDF (spray-drying – pasteurized fresh milk – E. cottonii).

3.5 Scoring value of parameters

Determination of the selected treatment was carried out by the scoring method on each observation criteria that has been carried out. This was done to show various considerations in determining the chosen treatment. Table 7 shows the scoring results of each observation criteria in the research which were based on the upper and lower mean value results obtained for each observation. Based on the table, it can be stated that raw material costs and process costs play a major role in determining the total scoring value. Based on Table 7, the treatment chosen was the spray-drying method – fresh milk inoculum media – E. cottonii encapsulation material (SDF) which has 22 as total score even though it was not treated with the best viability, EE, and added value.

Table 7

The scoring value of parameters observed on each treatment

Scoring value parameters FDS FDF SDS SDF Control
L. acidophilus (CFU/g) 4 3 3 3 0
Viability (%) 4 2 3 3 0
EE (%) 4 4 3 4 0
Added value (IDR/g) 4 4 3 4 1
Process cost 0 0 4 4 0
Raw material total cost 2 2 4 4 0
Total score 18 15 20 22 1

(a) FDS (freeze-drying – skim milk – E. cottonii), (b) FDF (freeze-drying – pasteurized fresh milk – E. cottonii), (c) SDS (spray-drying – skim milk – E. cottonii). (d) SDF (spray-drying – pasteurized fresh milk – E. cottonii).

3.6 Physicochemical, microbial, and structural characterization

The physicochemical characterization was carried out on the treatment with the highest score based on scoring results of final product observation criteria. The final product characteristics determine the product quality which was influenced by raw materials and the production process. The product morphological characterization and cell count of L. acidophilus for all treatments had been described in the previous section (Table 8).

Table 8

Characterization of spray-drying – pasteurized fresh milk with E. cottonii treatment

Properties Value
Moisture content % DB 15.78 ± 0.18
Water activity 0.294 ± 0.53
Hygroscopicity % 22.25 ± 0.11
Dissolution (min) 3.67 ± 0.82
In-vitro human gastrointestine
L. acidophilus (CFU/g) 4.77 ± 0.16
Population reduction (CFU/g) 4.95 ± 0.31

4 Discussion

Research on encapsulation methods, inoculum media, and coating materials has been carried out to find correct materials or process methods that can protect and control hydrated probiotic cells release in the gastrointestinal tract after consumption. The essence of this research was to know how the encapsulation method, inoculum media, and coating materials can affect the viability of L. acidophilus probiotic cells. The point was to find formulations and encapsulation methods that can maintain their viability and application in food products. The search for materials and hydration process method was also balanced with the added value analysis of hydrated L. acidophilus products. This was done as an experimental data simulation for scale-up preparation.

The criteria that must be considered in the scale-up production of hydrated L. acidophilus are viability, encapsulation efficiency, and production process duration. The production process duration will also determine the production cost which causes differences between value product, sales profits, and value-added. The encapsulation method with spray-drying allows it to be used as a method to produce large-scale hydrated L. acidophilus cells in a short time. The energy costs and the probiotic encapsulation process using the spray-drying method were also lower than the freeze-drying methods commonly used to dry or encapsulate the bioactive components. The drawback of the spray-drying method is the high temperature used (100–130°C) at both the inlet and outlet. Bacterial damage during spray-drying stems not only came from thermal effects, but also due to loss of water bound to the cell surface. One of the sites most prone to cell injury is the cytoplasmic membrane. The sequencing mechanism of cell viability decreasing due to cell damage by heat treatment was as follows [40]: (1) damage resulting from the drying media, (2) Aw of cell becomes low, and (3) heat induction occurs which causes the cell to have heat stress shock.

Encapsulation is the process of material coating by polymer or ceramic material so that it can protect the core material inside. The coating was carried out through a droplet of coating material on the core wall of the encapsulated material. Encapsulation can be mononuclear, poly-nuclear, and matrix. In this research, E. cottonii was used because it has hydrophilic properties. The advantages possessed by E. cottonii is that it can form a gel to trap probiotic cells, store cells in their gel cavity so that it can reduce the cell reduction population in the hydration process, food application, and in-vitro gastrointestinal testing [41]. The treatment using the FDS had the best viability value of 94.2%. The highest encapsulation efficiency was 11.84% with the treatment using FDF. This treatment had a cell viability value of 91.6% which was not significantly different from the treatment using SDS and SDF.

Based on the SEM test result, a significant difference between freeze-drying and spray-drying encapsulation results was the size of the hydrate particle produced. The size of the cell as a result of freeze-drying method was 1.8–11.4 µm with an encapsulation size of +22.13 µm (Figure 1), while the size of the cell as a result of spray-drying method was 3.8–5.5 µm with an encapsulation size of ±17.97 µm. The particle size of the spray-drying cell encapsulation is smaller than the result of freeze-drying, because in the spray-drying process, the particles are passed through the nozzle so that the resulting particle size is smaller and uniform [27]. The inoculum media also protects the hydrated cell particles. Skim milk provides a layer that hardens the encapsulation wall, while fresh milk provides a soft layer that appears to fill the surface of the cell encapsulation wall. The fat content in fresh milk can protect by blocking moisture transport due to its relatively low polarity [42].

The E. cottonii coating material forms bubbles that trap more cells in the sac. The cells would enter the cavity or sac formed by E. cottonii to provide a bone texture on the cells’ surface [43]. E. cottonii was able to prevent hydrated cell surface cracking by strengthening the cell wall structure. The function of E. cottonii in the hydrate capsule provides a framework that strengthens the cell capsule [41]; however, the viability of the encapsulation results was not only influenced by the composition of the coating material or the inoculum medium. Process duration can also play a major role in decreasing the viability of the encapsulated cells [44]. The reinforced structure of E. cottonii had an unfavorable effect on the combination of skim milk, because its structure was bony, causing cells to appear damaged. In contrast, the combination of E. cottonii with fresh milk inoculum media, the void in the capsule appears to be covered by a bubbling fat structure that eventually forms an elongated capsule structure.

The added value calculated in this research was not influenced by costs arising from the packaging, shipping, and marketing processes. Uswatun and Mayshuri (2015) [45] reported that the calculation of added value based on the Hayami and Kawagoe method only focuses on calculating the added value of the initial commodity and the production process. The advantage of using the Hayami and Kawagoe method is that the owners of the capital can focus more on the value of raw materials and the production processes that were carried out to increase their added value. Sindhu et al. (2019) [46] reported that the Hayami and Kawagoe method can further help owners of production capital to decide on raw materials and production processes that will be used in the production of the goods they want.

Based on the observation ranking value on added value, value-added ratio, labor share, profit, profit level, profit margin, and company owner profit depicted in Table 7, the best treatment was using SDF. This treatment was also the best treatment from the scoring method on each observation criteria with an overall score value of 22. Treatment using SDF has the lowest raw material cost and had a process cost that did not significantly differ from the lowest processing cost.

Based on the research results, it can be concluded that fresh milk can be used as an inoculum media to reduce raw material costs. In spray-drying method, the viability value of the hydrated L. acidophilus cells produced through growth in fresh milk inoculum media was not significantly different from skim milk inoculum media growth. Based on statistical tests, the use of E. cottonii did not have a significant effect on the effectiveness of encapsulation. E. cottonii provides a bony structure that can protect cell walls from damage due to the encapsulation process. The drying method has a significantly different effect on the number of L. acidophilus cells contained in dry cells and also affects its viability. The final decision should be left to the company capital owners in developing an industrial scale. Choosing the treatment with the best added value or the largest number of probiotic cells, even though overall SDF treatment already has the criteria as a candidate probiotic preparation, namely having a minimum number of probiotic cells of 107 CFU/g before consumption.

The research results can be used as consideration for production factor owners to assess the feasibility of the production process and raw materials to be used in producing hydrated L. acidophilus. Based on the research results, spray-drying can provide a greater value add of 10–20% than the freeze-drying method.

5 Conclusion

The value-added investigation plays a vital part in deciding the foremost appropriate downstream strategy. The spray-drying method in the L. acidophilus cell encapsulation can reduce the production cost because it can produce more output in a shorter time than the freeze-drying method. The process cost reduction can increase the added value of the cell encapsulation produced by this method. The technology used can produce more output and further reduce labor share and increase the profits for the production capital owners. In addition to the production process, raw materials processing also plays role in the value-added calculation. This research gives information in determining the choice of production processes, and encapsulation materials are not only the physicochemical and microbiological properties that need to be considered but also the added value of the product which determines the profitability of the production factors owner.

Acknowledgments

The authors thank the Laboratory Head and Laboratory Assistant of Food Microbiology Laboratory, Food Chemistry Laboratory, and Food Engineering Laboratory of Food Processing Industry Technology. Faculty of Agricultural Industrial Technology department, Padjadjaran University Jatinangor and Chemistry Instrument Laboratory of Mathematics, Science and nature faculty, Indonesian Education University where the research was conducted.

  1. Funding information: The authors state no funding involved.

  2. Author contributions: S.Y.: conceptualization, data curation, formal analysis, funding acquisition, investigation, methodology, project administration, resources, software, and writing the original draft. R.K.: conceptualization, supervision, writing review and editing, and validation. E.S.: conceptualization, supervision, and writing review and editing. E.M.: conceptualization, supervision, and writing review and editing.

  3. Conflicts of interest: The authors state no conflict of interest.

  4. Data availability statement: All data generated or analyzed during this study are included in this published article.

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Received: 2020-12-30
Revised: 2022-03-14
Accepted: 2022-03-15
Published Online: 2022-04-27

© 2022 Silvia Oktavia Nur Yudiastuti et al., published by De Gruyter

This work is licensed under the Creative Commons Attribution 4.0 International License.

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https://doi.org/10.1515/opag-2022-0081
Schlagwörter für diesen Artikel
formulation; Hayami and Kawagoe; hydrated cell; process cost; techno-economic analysis
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  32. Strategies for overcoming farmers’ lives in volcano-prone areas: A case study in Mount Semeru, Indonesia
  33. Principal component and cluster analyses based characterization of maize fields in southern central Rift Valley of Ethiopia
  34. Profitability and financial performance of European Union farms: An analysis at both regional and national levels
  35. Analysis of trends and variability of climatic parameters in Teff growing belts of Ethiopia
  36. Farmers’ food security in the volcanic area: A case in Mount Merapi, Indonesia
  37. Strategy to improve the sustainability of “porang” (Amorphophallus muelleri Blume) farming in support of the triple export movement policy in Indonesia
  38. Agrarian contracts, relations between agents, and perception on energy crops in the sugarcane supply chain: The Peruvian case
  39. Factors influencing the adoption of conservation agriculture by smallholder farmers in KwaZulu-Natal, South Africa
  40. Meta-analysis of zinc feed additive on enhancement of semen quality, fertility and hatchability performance in breeder chickens
  41. Meta-analysis of the potential of dietary Bacillus spp. in improving growth performance traits in broiler chickens
  42. Biocomposites from agricultural wastes and mycelia of a local mushroom, Lentinus squarrosulus (Mont.) Singer
  43. Cross transferability of barley nuclear SSRs to pearl millet genome provides new molecular tools for genetic analyses and marker assisted selection
  44. Detection of encapsulant addition in butterfly-pea (Clitoria ternatea L.) extract powder using visible–near-infrared spectroscopy and chemometrics analysis
  45. The willingness of farmers to preserve sustainable food agricultural land in Yogyakarta, Indonesia
  46. Transparent conductive far-infrared radiative film based on polyvinyl alcohol with carbon fiber apply in agriculture greenhouse
  47. Grain yield stability of black soybean lines across three agroecosystems in West Java, Indonesia
  48. Forms of land access in the sugarcane agroindustry: A comparison of Brazilian and Peruvian cases
  49. Assessment of the factors contributing to the lack of agricultural mechanization in Jiroft, Iran
  50. Do poor farmers have entrepreneurship skill, intention, and competence? Lessons from transmigration program in rural Gorontalo Province, Indonesia
  51. Communication networks used by smallholder livestock farmers during disease outbreaks: Case study in the Free State, South Africa
  52. Sustainability of Arabica coffee business in West Java, Indonesia: A multidimensional scaling approach
  53. Farmers’ perspectives on the adoption of smart farming technology to support food farming in Aceh Province, Indonesia
  54. Rice yield grown in different fertilizer combination and planting methods: Case study in Buru Island, Indonesia
  55. Paclobutrazol and benzylaminopurine improve potato yield grown under high temperatures in lowland and medium land
  56. Agricultural sciences publication activity in Russia and the impact of the national project “Science.” A bibliometric analysis
  57. Storage conditions and postharvest practices lead to aflatoxin contamination in maize in two counties (Makueni and Baringo) in Kenya
  58. Relationship of potato yield and factors of influence on the background of herbological protection
  59. Biology and life cycle Of Diatraea busckella (Lepidoptera: Crambidae) under simulated altitudinal profile in controlled conditions
  60. Evaluation of combustion characteristics performances and emissions of a diesel engine using diesel and biodiesel fuel blends containing graphene oxide nanoparticles
  61. Effect of various varieties and dosage of potassium fertilizer on growth, yield, and quality of red chili (Capsicum annuum L.)
  62. Review Articles
  63. Germination ecology of three Asteraceae annuals Arctotis hirsuta, Oncosiphon suffruticosum, and Cotula duckittiae in the winter-rainfall region of South Africa: A review
  64. Animal waste antibiotic residues and resistance genes: A review
  65. A brief and comprehensive history of the development and use of feed analysis: A review
  66. The evolving state of food security in Nigeria amidst the COVID-19 pandemic – A review
  67. Short Communication
  68. Response of cannabidiol hemp (Cannabis sativa L.) varieties grown in the southeastern United States to nitrogen fertilization
  69. Special Issue on the International Conference on Multidisciplinary Research – Agrarian Sciences
  70. Special issue on the International Conference on Multidisciplinary Research – Agrarian Sciences: Message from the editor
  71. Maritime pine land use environmental impact evolution in the context of life cycle assessment
  72. Influence of different parameters on the characteristics of hazelnut (var. Grada de Viseu) grown in Portugal
  73. Organic food consumption and eating habit in Morocco, Algeria, and Tunisia during the COVID-19 pandemic lockdown
  74. Customer knowledge and behavior on the use of food refrigerated display cabinets: A Portuguese case
  75. Perceptions and knowledge regarding quality and safety of plastic materials used for food packaging
  76. Understanding the role of media and food labels to disseminate food related information in Lebanon
  77. Liquefaction and chemical composition of walnut shells
  78. Validation of an analytical methodology to determine humic substances using low-volume toxic reagents
  79. Special Issue on the International Conference on Agribusiness and Rural Development – IConARD 2020
  80. Behavioral response of breeder toward development program of Ongole crossbred cattle in Yogyakarta Special Region, Indonesia
  81. Special Issue on the 2nd ICSARD 2020
  82. Perceived attributes driving the adoption of system of rice intensification: The Indonesian farmers’ view
  83. Value-added analysis of Lactobacillus acidophilus cell encapsulation using Eucheuma cottonii by freeze-drying and spray-drying
  84. Investigating the elicited emotion of single-origin chocolate towards sustainable chocolate production in Indonesia
  85. Temperature and duration of vernalization effect on the vegetative growth of garlic (Allium sativum L.) clones in Indonesia
  86. Special Issue on Agriculture, Climate Change, Information Technology, Food and Animal (ACIFAS 2020)
  87. Prediction model for agro-tourism development using adaptive neuro-fuzzy inference system method
  88. Special Issue of International Web Conference on Food Choice and Eating Motivation
  89. Can ingredients and information interventions affect the hedonic level and (emo-sensory) perceptions of the milk chocolate and cocoa drink’s consumers?
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Heruntergeladen am 20.12.2025 von https://www.degruyterbrill.com/document/doi/10.1515/opag-2022-0081/html
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