Biosurfactants and Bioemulsifiers

Dr Pattanathu Rahman

Biotechnology Programme Leader

Teesside University - UK

&

Founding Director

TeeGene Biotech Ltd.

Wilton Centre, Wilton – UK

http://www.teegene.co.uk

TeeGene Biotech Ltd.TeeGene Biotech Ltd.

TEESSIDE UNIVERSITY - UK

Wilton Centre http://www.wiltoncentre.com/

• Reg. No: 09361483

• Registered Office: ONE AMERICA SQUARE, CROSSWALL, LONDON - EC3N 2SG

• Office: H245, WILTON CENTRE, REDCAR – TS104RF

CPINHCNBMCFujifilmFORGETeeGene

BIOSURFACTANTS

Properties:

� Non-toxic, Non-hazardous

� Thermostable

� Low surface tension

� Emulsify: wide range of hydrocarbons - Stable emulsion

• Teegene biotech is a spin out venture from Teesside University, which is pioneering the use of biosurfactants in a range of cosmetics, environmental and biomedical applications.

• Highly-effective biotechnology based surface-active multi-functional products

Classification of microbial surfactants

Microbial surfactants

Biosurfactants Bioemulsifier

• Low molecular weight• Composed of sugars,

amino acids and fatty acids

• Reduce surface/interfacial tensions

• Solubilise and emulsify hydrophobic substrates

• High molecular weight• Consists of

polysaccharide-protein lipid complex

• Less effective in surface/interfacial tension reduction

• Efficient emulsification and stabilization of emulsion

Characteristics• Both bioemulsifiers and biosurfactants contain hydrophobic and

hydrophilic moieties

• The non-polar (hydrophobic) region is often a hydrocarbon chain.

• The polar portion (hydrophilic) region may be ionic (cationic or anionic), non-ionic or amphoteric

(Campos et al., 2013)

Chemical structures of the conventional mannosylerythritol lipids.

(Fukuoka 2008)

Surfactin (Mulligan, 2009)

(Zhou et al.,2013)

Sophorolipid

Structure of Biosurfactants

Marine Microbes

Harvey and Rahman, 2015

Culture conditions required for Algae

Harvey and Rahman, 2015

Screening of novel microbes from nature

Biosurfactant Bacteria - Teesside

AM419154 Pseudomonas teessidea partial 16S rRNA gene, type strain PR65T

AM419155 Pseudomonas clemancea partial 16S rRNA gene, type strain PR221T

Culture Collection = 122 bacterial strains from Northeast England

Scale up of Bioprocess

� 100ml of broth (Nutrient: Glycerol + MSM)

� Conditions: agitation 150rpm, pH 7.0 and incubation at 30oC

Biosurfactant production in batch fermenter (New Brunswick Bioflo 101)

HPLC / Mass spectrum of rhamnolipids

Mass (m/z)

503.654 g/mol

649.595 g/molS#: 3-62 RT: 0.04-1.00 AV: 60 NL: 2.82E7

F: - c ms [ 200.00 - 800.00]

200 250 300 350 400 450 500 550 600 650 700 750 800

m/z

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90

95

100

Rela

tive A

bundance

649.5

650.4

503.7

746.9

677.2

762.9648.8

678.3505.0 763.8258.6 502.5

479.3333.3270.7 529.4 621.5 684.6242.7

339.5 703.2601.3573.5451.4367.1 417.4

Emulsification of hydrocarbons by biosurfactants

OIL EXTRACTION TECHNIQUE

Tertiary Recovery or Enhanced Oil Recovery (EOR)

•MEOR is a method of tertiary recovery •EOR could potentially recover a further 60 billion barrels (US Report) •Current oil extraction techniques still leave oil trapped in low permeability zones•MEOR can potentially solve this problem. •MEOR is a ‘green’ option to extract remaining reservoir oil

Sandstone

Housing Oil

Shale

Shale

Overburden

Hot Water

Steam Injector

Steam

Oil Reservoir Tertiary Recovery (Collitt, Sina and Rahman 2014)

BEOR TECHNOLOGY

http://www.huffingtonpost.com/amanda-greene/soy-lecithin-why-is-it-in_b_2891780.html

BIOSURFACTANT MARKET RESEARCH

�According to F&S estimations the global biosurfactants market rounds $1.76 B (base year 2013), with a combined annual growth rate of about 3.5%

�The global biosurfactant market is expected to reach $2.8 B in 2023

�European markets are leading the biosurfactant space with 53.8% of global sales revenue, followed by North America and then Asia Pacific.

�(Source: Life Sciences and Biotech Technical Insight, Frost & Sullivan, 22 May 2014)

SEKHON, K.K. AND RAHMAN, P.K.S.M. 2014. Frontiers in Microbiology 5:454-460.

ALGAL EXOPOLYSACCHARIDES

• Algal EPS represent a huge range of structures

• They are high-molecular-weight structures (10-30 kDa)

• EPS structure

• varies widely between different genera of algae

• environmental conditions

(Patel et al., 2010)

POLYSACHHARIDES - BIOEMULSIFIERS

• Worldwide, production of polysaccharides from marine biomass is between 25,000-30,000 tons/year (Pichavant, 2009)

• polysaccharides can be classified in to three groups: 1. Cytosolic polysaccharides – provide carbon and energy

sources for the cell,

2. Structural polysaccharides - make up the cell wall

3. Exopolysaccharides (EPS) - secreted in to the extracellular environment in capsules or biofilms

(Roger 2002)

SCREENING OF ALGAE

A. Saltburn, B. Seaton and C. Skinningrove beaches

(Google Maps, 2015).Harvey and Rahman, 2015

Surface tension measurements Emulsification Index (E24 %) in

Sunflower Oil

Harvey and Rahman, 2015

Bacterial Bio-emulsifier

Algal bio-emulsifiers

Harvey and Rahman, 2015

TECHNOLOGY COLLABORATION WITH NETWORK MEMBERS

Green micro-algae Dunaliella salina and red algae Porphyridium cruentum –bioemulsifier producers

ANY SLIMY ALGAE:

Preliminary screening for biosurfactants and bioemulsifiers - Free of cost

Contact me (rahman@teegene.co.uk)

ACKNOWLEDGEMENT

� Teesside University

� Commonwealth Scholarship Commission

� Business Investment Funds

� Biobase Europe Pilot Plant (Belgium)

� Research and Development Team

� BSc, MEng, MSc and PhD students

Thanks

Dr Pattanathu Rahman

Biotechnology Programme Leader

Teesside University - UK

&

Founding Director

TeeGene Biotech Ltd.

Wilton Centre, Wilton – UK

Email: rahman@teegene.co.uk