MANNOSYLERYTHRITOL LIPIDS (MEL) AS ADDITIVES IN COSMETIC FORMULATIONS

Transcription

1 MANNOSYLERYTHRITOL LIPIDS (MEL) AS ADDITIVES IN COSMETIC FORMULATIONS Fraunhofer Institute for Interfacial Engineering and Biotechnology IGB Alexander Beck SuperBIO Workshop Biosurfactants, Gent, Belgium User Group Meeting Biosurfactants Carbosurf, 3rd April 2017

2 ...the next promising biosurfactants produced by microorganisms Literature available for lab scale synthesis and applications Large gap towards process optimization and piloting scale Addressed in the Carbosurf Project as one of four product lines together with Sophorolipids, Rhamnolipids and Xylolipids Microbial Bios urfactants Origin Properties and Potential Applications Titers Mannosylerythritol lipids (MEL) Cellobiose lipids (CL) Pseudozyma sp. Self assembling properties Emulsifier Cosmetics and personal care Pharmaceuticals Cleaning agents Ustilago sp. Antimicrobial effects Preservatives Cleaners 160 g/l 33 g/l

3 Current research on MEL production at IGB Strain selection Evaluation of renewable resources Metabolic pathway analysis Bioprocess engineering Medium composition Reactor design Process strategies Controllability Downstream processing Aim High space-time-yield Scale-up engineering

4 Chemical structure of Mannosylerythritol Lipids Glycolipids contain hydrophilic sugar moiety and fatty acid residues Microorganisms produce a mixture of different MEL structures R 6 = Ac- or H- C2 - C18 R 4 = Ac- or H- C2 - C18

5 polarity Different MEL variants lead to different properties MELs clustered according to polarity on TLC (Kitamoto et al., 1990) Oil Fatty acid MEL-A MEL-B MEL-C MEL-D MEL-B MEL-A MEL-C TLC result MEL-D Kitamoto, D., Akiba, S., Hioki, C., Tabuchi, T., (1990) Extracellular Accumulation of Mannosylerythritol Lipids by a Strain of Candida antarctica. Agricultural and Biological Chemistry 54,

6 How to tailor MEL production? Different approaches possible (simultaneously): Use of different strains Use of different (renewable) feedstocks Post-modification of produced MEL-mixture Modification of metabolic pathway

7 polarity Selection of production strain Use of different strains (Morita et al., 2008) Morita, T., Konishi, M., Fukuoka, T., Imura, T., Kitamoto, D., (2008) Production of glycolipid biosurfactants, mannosylerythritol lipids, by Pseudozyma siamensis CBS 9960 and their interfacial properties. Journal of bioscience and bioengineering 105,

8 Selection of production strain Pseudozyma strain 2 produces all main MEL derivatives and a high titer of hydrophilic MEL-D P. strain 3 produces mainly MEL-B and MEL-C Different Pseudozyma strains

9 Variation of MEL composition induced by carbon source Use of different (renewable) feedstocks Hydrophilic: Molasses, Lignocellulosic sugars, Glycerol, Used for biomass growth Hydrophobic: Soybean oil, Olive oil, Coconut oil, Castor oil, Necessary for MEL synthesis Influences side-chain composition Oleic acid Ricinoleic acid Lauric acid Different oils

10 polarity Post-modification of produced MEL-mixture Enzymatic treatment with lipase at 60 C Increases percentage of hydrophilic MEL variants MEL-D Mono-acylated MEL-X

11 How to set up a scalable fermentation process? Strategies for a controllable fermentation process Metabolism Operating Conditions Media Components C-Sources Genetics / Biosynthesis Stirring, Foam, O 2 Supply Mineral Media Oil, Fatty Acids, Hydrophilic C Expression Level of Target Genes Regulatory Controls Repression (C or N) Inhibition Induction Overflow Metabolites Antifoam/ Defoamers Feeding Strategies Aeration and Stirring Strategy ph Control Seed Culture Seed Culture Media C- and N-source Amino Acids Vitamins Trace Elements Un-/saturated Chain Length (C12-C18) Water-soluble C- Sources: Glycerol, Glucose

12 How to set up a scalable fermentation process? Strategies for a controllable fermentation process Repeated-Batch strategy with oil addition after growth phase Control of ph, stirring and po 2 Growth phase Production phase Growth phase Production phase Repeated oil addition

13 Physico-chemical properties of MEL Measurement of surfactant properties Strain Substrate (oils) CMC [m g/l] σ min [mn/m] Oil A Oil B 2 25 Pseudozyma 2 Oil C Oil D Oil E Pseudozyma 1 Oil A Different foaming ability of MEL structures

14 Application of MEL in cosmetics Reported potential applications in literature: Moisturizing effect on dry skin (ceramide-like properties of MEL-A) Repair of damaged hair (MEL-A and B) Activation of fibroblasts and papilla cells Antioxidant activities anti-aging ingredients? Work performed by Croda: High throughput screening for formulations in personal care Extended functionalities Currently fermentation running to supply samples Morita, T., Fukuoka, T., Imura, T., Kitamoto, D., (2013) Production of mannosylerythritol lipids and their application in cosmetics. Appl Microbiol Biotechnol 97,

15 Summary and Outlook Great structural variety of MELs Tailoring of MEL variants already possible Development of a scalable production process Properties of MEL make them suitable for cosmetic applications

16 Acknowledgement Key researchers: Dr. Michael Günther Paula Carrillo Head of Department: apl. Prof. Dr. Steffen Rupp Contact Alexander Beck, M.Sc. Dr. Susanne Zibek Nobelstrasse 12, Stuttgart, Germany