Startseite Production of Non-Toxic Biosurfactant – Surfactin – Through Microbial Fermentation of Biomass Hydrolysates for Industrial and Environmental Applications
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Production of Non-Toxic Biosurfactant – Surfactin – Through Microbial Fermentation of Biomass Hydrolysates for Industrial and Environmental Applications

  • Buddhi P. Lamsal , Pathra Patra , Rajat Sharma und Christopher C. Green
Veröffentlicht/Copyright: 1. Oktober 2019
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Tenside Surfactants Detergents
Aus der Zeitschrift Tenside Surfactants Detergents Band 56 Heft 5

Abstract

The economically viable large-scale production of the pure isoforms of the surfactin biosurfactants, involving bacterial – Bacillus subtilis – fermentation of biomass hydrolysate feedstock, relies on the types of bacterial strains, optimization of the fermentation processing parameters, differences in the composition of the carbon and nitrogen in the bacterial media, and the chromatography techniques used for isolation of the isoforms. Here, we biosynthesized the surfactin isoforms in their mixture forms through fermentation of biomass hydrolysates at 2 wt.% carbohydrate content. The surfactin isoforms were assessed for their surface-active properties and toxicity. The enzyme hydrolysates considered were from switchgrass, soyhull (fiber), alfalfa, and bagasse. The isoform mixtures obtained after fermentation of the hydrolysates and, glucose as a control, were concentrated using chromatography columns, and characterized for molecular weights (MWs) and relative distribution using LCMS. The isoform mixtures, obtained in different fermenters (5- and 15-L) and, for different hydrolysates, invariably constituted 5 isoforms with MWs as 992.6, 1006.6, 1020.6, 1034.6, 1048.6, 1062.6 m/z amu, with their relative proportions as 6, 24, 35, 24, and 10 weight % respectively. The surface tension values of all these isoforms, in the absence of electrolytes and at 12 ppt salinity, were similar: 37 (pH 6.5) and 31 (pH 9.5) mN/m. Furthermore, the emulsification index values for the isoforms were also similar: Dispersant-to-Oil ratio as 1:20. The LC50 for Gulf killifish, Fundulus grandis for these surfactin isoforms ranged between 10 and 20 mg/L; a microbially-produced surfactin variant FA-Glu (Fatty acid Glutamate) was least toxic with LC50 at ∼100 mg/L. Thus, the surfactin synthesis approach adopted here suggested that pure (>95 wt.%) non-toxic isoforms of surfactin biosurfactants can be produced in the forms of their mixtures with surface-active properties similar to those of the pure forms of the surfactin isoforms.

Kurzfassung

Die wirtschaftlich sinnvolle großtechnische Herstellung der reinen Isoformen des Surfactins bei der Fermentation des Biomasse-Hydrolysat-Ausgangsmaterials mit Bacillus subtilis beruht auf den Bakterienstämmen, der Optimierung der Fermentationsparameter, den Unterschieden in der Zusammensetzung von Kohlenstoff und Stickstoff in den Bakterienmedien und die zur Isolierung der Isoformen verwendeten Chromatographietechniken. In dieser Arbeit biosynthetisierten wir die Surfactin-Isoformen in ihren Mischungen durch Fermentation von Biomasse-Hydrolysaten bei einem Kohlenhydratgehalt von 2 Gew.-%. Die Surfactin-Isoformen wurden hinsichtlich ihrer oberflächenaktiven Eigenschaften und Toxizität bewertet. Die untersuchten Enzymhydrolysate stammten aus Rutenhirse, Sojaschale (Ballaststoffe), Luzerne und Bagasse. Die nach Fermentation der Hydrolysate und Glucose (als Kontrolle) erhaltenen Isoformenmischungen wurden in Chromatographiesäulen konzentriert und ihre Molekulargewichte (MWs) und relative Verteilung mit der LCMS zur Charakterisierung bestimmt. Die Isoformenmischungen, die in verschiedenen Fermentern (5 L und 15 L) für verschiedene Hydrolysate erhalten wurden, bestanden ausnahmslos aus 5 Isoformen mit einem Molekulargewicht von 992,6, 1006,6, 1020,6, 1034,6, 1048,6 und 1062,6 m/z amu. Ihre relativen Anteile waren 6, 24, 35, 24 bzw. 10 Gew.-%. Die Oberflächenspannungen aller dieser Isoformen waren in Abwesenheit von Elektrolyten und bei einem Salzgehalt von 12 ppt ähnlich: 37 mN/m (pH 6,5) und 31 mN/m (pH 9,5). Darüber hinaus waren auch die Emulgierungsindices für die Isoformen ähnlich: Dispergiermittel-zu-Öl-Verhältnis 1:20. Die LC50-Werte der Surfactin-Isoformen (bestimmt für Golfkillifische (Fundulus grandis) lagen zwischen 10 mg/L und 20 mg/L. Die mikrobiell erzeugte Surfactin-Variante FA-Glu (Fettsäure-Glutamat) war mit einem LC50-Wert von∼100 mg/L am wenigsten toxisch. Der hier angewandte Ansatz zur Surfactinsynthese zeigt somit deutlich, dass reine (>95 Gew.-%) nichttoxische Surfactinisoformen in ihren Mischungen hergestellt werden können, deren oberflächenaktiven Eigenschaften denen der reinen Surfactin-Isoformen ähnlich sind.

Key words: Surfactin isoforms; pretreatment; fermentation surface tension; marine toxicity
Stichwörter: Surfactin-Isoformen; Vorbehandlung; Oberflächenspannung bei der Fermentation; Meerestoxizität
∗Correspondence address, Prof. Dr. Buddhi P. Lamsa, Department of Food Science and Human Nutrition, Iowa State University, 2312 Science Building, Iowa State University, Ames, IA 20010, USA, Tel.: +1 (515)294-8681, E-Mail:

Dr. Buddhi Lamsal completed his postdoctoral research in food science at Iowa State University in 2006 after completing a Ph.D. from the University of Wisconsin-Madison in 2004. He was a Research Assistant Professor at Kansas State University from 2006–2008. He started to work as an Assistant Professor in 2008 and became an Associate Professor in 2014 in Food Engineering and Bioprocessing at the Department of Food Science and Human Nutrition at Iowa State University. His research includes the processing of food, ingredients-function relationship, and bioprocessing fermentation for biobased products.

Dr. Partha Patra earned his doctorate degree in Materials Engineering from Indian Institute of Science, Bangalore. His research efforts focus on dealing with challenges associated with earth resource recovery and management, and in developing interfacial engineering tools to promote the application of bioreagents in personal care, pharmaceuticals, and food industries.

Mr. Rajat Sharma is a Ph.D. candidate in Food Science and Human Nutrition department at Iowa State University since 2012. He started his research on the utilization of renewable biomass sugars for microbial surfactant production. He earned a masters in Biological and Agricultural Engineering from North Carolina State University in 2012. Rajat since 2017 has also been working as a Sr. Production Engineer at Valent Biosciences in Osage Iowa, where his work is focused on managing large scale bacterial fermentation for bio-rational products for the agro and public health industry.

Dr. Christopher Green is an Associate Professor at LSU AgCenter School of Renewable Natural Resources. After completing a Ph.D. in 2007 from Southern Illinois University, Carbondale in Physiology, Dr. Green joined the LSU AgCenter faculty in 2008. Dr. Green's lab is centered within the field of applied reproductive fish physiology and endocrinology, with a focus on freshwater and estuarine aquaculture species.

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Received: 2019-06-16
Accepted: 2019-07-24
Published Online: 2019-10-01
Published in Print: 2019-09-16

© 2019, Carl Hanser Publisher, Munich

Artikel in diesem Heft

  1. Contents/Inhalt
  2. Contents
  3. Microbial Synthesis
  4. Production of Non-Toxic Biosurfactant – Surfactin – Through Microbial Fermentation of Biomass Hydrolysates for Industrial and Environmental Applications
  5. Characterisation Novel Biosurfactants
  6. Structures and Properties of Sophorolipids in Dependence of Microbial Strain, Lipid Substrate and Post-Modification
  7. Personal Care/Cleansing
  8. Amino-Acid Surfactants in Personal Cleansing (Review)
  9. Toward Milder Personal Care Cleansing Products: Fast ex vivo Screening of Irritating Effects of Surfactants on Skin Using Raman Microscopy
  10. Textile Surface Modification
  11. Surface Characterization of Textiles for Optimization of Functional Polymeric Nano-Capsule Attachment
  12. Enhanced Oil Recovery and Oil-Spill Dispersants
  13. Pseudo-Gemini Biosurfactants with CO2 Switchability for Enhanced Oil Recovery (EOR)
  14. Hydrophilic-Lipophilic-Difference (HLD) Guided Formulation of Oil Spill Dispersants with Biobased Surfactants
  15. Mineral Processing
  16. Floatability of Chalcopyrite by Glycolipid Biosurfactants as Compared to Traditional Thiol Surfactants
  17. Antimicrobial Properties
  18. Stability of Emulsions and Nanoemulsions Stabilized with Biosurfactants, and their Antimicrobial Performance against Escherichia coli O157:H7 and Listeria monocytogenes
https://doi.org/10.3139/113.110644
Schlagwörter für diesen Artikel
Surfactin isoforms; pretreatment; fermentation surface tension; marine toxicity

Artikel in diesem Heft

  1. Contents/Inhalt
  2. Contents
  3. Microbial Synthesis
  4. Production of Non-Toxic Biosurfactant – Surfactin – Through Microbial Fermentation of Biomass Hydrolysates for Industrial and Environmental Applications
  5. Characterisation Novel Biosurfactants
  6. Structures and Properties of Sophorolipids in Dependence of Microbial Strain, Lipid Substrate and Post-Modification
  7. Personal Care/Cleansing
  8. Amino-Acid Surfactants in Personal Cleansing (Review)
  9. Toward Milder Personal Care Cleansing Products: Fast ex vivo Screening of Irritating Effects of Surfactants on Skin Using Raman Microscopy
  10. Textile Surface Modification
  11. Surface Characterization of Textiles for Optimization of Functional Polymeric Nano-Capsule Attachment
  12. Enhanced Oil Recovery and Oil-Spill Dispersants
  13. Pseudo-Gemini Biosurfactants with CO2 Switchability for Enhanced Oil Recovery (EOR)
  14. Hydrophilic-Lipophilic-Difference (HLD) Guided Formulation of Oil Spill Dispersants with Biobased Surfactants
  15. Mineral Processing
  16. Floatability of Chalcopyrite by Glycolipid Biosurfactants as Compared to Traditional Thiol Surfactants
  17. Antimicrobial Properties
  18. Stability of Emulsions and Nanoemulsions Stabilized with Biosurfactants, and their Antimicrobial Performance against Escherichia coli O157:H7 and Listeria monocytogenes
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