USE OF FUNGAL/PLANT ENZYMES IN BIOTECHNOLOGY

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Contents• Introduction to enzymes Fungal/plant enzyme• Introduction to biotechnology• Use of fungal/plant enzymes in biotechnology• In food industry• In baking• In animal feed• In beverages• In detergents• In pharmaceutical• In textile

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• In pulp and paper industry• In organic synthesis industry• Recent research work done in PakistanConclusionReferences

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Enzymes are biological catalysts or assistants. Enzymes consist of various types of proteins that work to drive the chemical reaction required for a specific action or nutrient .

Introduction to Enzymes

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key roles in numerous biotechnology products and processes that are commonly encountered in the form of food and beverages, cleaning supplies, clothing, paper products, transportation fuels, pharmaceuticals, and monitoring devices.

Industrial enzyme market has expanded at a rate of about 10% annually, microbial enzymes have largely replaced the traditional plant and animal enzymes, and most of them are produced recombinantly.

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Classification of enzymes:According to the Enzyme Commission, International Union of Biochemistry and Molecular Biology enzymes are classified into following.

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How Enzymes Work??

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Biotechnology: Biotechnology is defined as the exploitation of biological processes for industrial and other purposes, Biotechnology can be broadly defined as "using organisms or their products for commercial purposes."especially the genetic manipulation of micro organisms for the production of antibiotics, hormones, etc

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Role of fungal/plant enzymes in biotechnologyRole of fungal/plant enzymes in biotechnology

Fungi have vital roles in biotechnology such as production of drugs and enzymes (Archer et al., 2008).

Fungi can be cultured easily and hence they can be used in microbiological, genetic and molecular research (Hoffmeister and Keller, 2007).

Aspergillus is wide spread fungus in nature, including soil, colonizing plant materials and decomposing agricultural crops (Varga et al., 2004) as well as most common air borne fungi (Gregory, 1973). Raghukumar C. 2008

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fungi thrive in diverse environments and can exploit marginal living conditions in large part because they produce different enzymes including laccases, cellulases, catalases and superoxide dismutases which are capable of performing difficult chemical reactions (Wheeldon et al., 2008).

Archer, (2000) described that total sixteen (16) fungal enzymes are used in the food industry and thirteen (13) of them has been obtained from Aspergillus.

Photo by: Fyle

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Biotechnology

Pharmaceutical industry

Food and beverages

Paper and pulp industry

Biofuel industry

Other organic synthesis

Textile

Biotechnology encompasses wide range of industries such as

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Use of fungal/plant enzymes in starch and fuel industry

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Industry Enzyme class Application References

Starch and fuel Amylase Starch liquefaction and saccharification

C.A. Voigt, S. Kauffman, Z.G. Wang,2000

Amyloglucosidase SaccharificationT. Godfrey, S.I. West, (1996)

Pullulanase SaccharificationC.A. Voigt, S. Kauffman, Z.G. Wang, (2000)

Glucose isomerase Glucose to fructose conversion

M.M. Altamirano, J.M. Blackburn, C. Aguayo, A.R. Fersht, (2000)

Cyclodextrin-glycosyltransferase Cyclodextrin production

T. Godfrey, S.I. West, (1996)

Xylanase Viscosity reduction (fuel and starch)

Andrade SV, Polizeli MLTM, Terenzi HF, Jorge JA (2004)

Protease Protease (yeast nutrition – fuel)

Fuglsang et al., (1995)

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In December 2014, Novozymes launched first ever enzymatic process using a liquid lipase – Eversa® Transform – for the production of biodiesel. in Biodiesel Magazine,February 2015

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Use of fungal/plant enzymes in food industry

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Industry Enzyme class Application References

Food (including dairy) Protease Milk clotting, infant formulas (low allergenic), flavor

P.L. Wintrode, K. Miyazaki, F.H. Arnol, (2000)

Lipase Cheese flavor•Karl-Erich Jaeger•Manfred T Reetz, (1998)

Lactase Lactose removal (milk)M.W. Christensen, L. Andersen, O. Kirk, H.C. Holm, (2001)

Pectin methyl esterase Firming fruit-based productsGalante et al. 1998b, Godfrey and West 1996b and Uhlig, (1998)

Pectinase Fruit-based productsGalante et al. 1998b, Godfrey and West 1996b and Uhlig, (1998)

Transglutaminase Modify visco-elastic propertieGrassin and Fauquembergue 1996b and Uhlig , (1998)

Acid and thermostable pectinase

Improvement in pressing fruit mashes and high colour extraction

Grassin and Fauquembergue 1996b and Uhlig 1998

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Use of fungal/plant enzymes in baking industry

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Industry Enzyme class Application References

Baking Amylase Bread softness and volume, flour adjustment

M.B. Tobin, C. Gustafsson, G.W. Huisman, (2000)

Xylanase Dough conditioningAndrade SV, Polizeli MLTM, Terenzi HF, Jorge JA (2004)

LipaseDough stability and conditioning (in situ emulsifier)

•Karl-Erich Jaeger•Manfred T Reet, (1998)

PhospholipaseDough stability and conditioning (in situ emulsifier)

K. Clausen, (2001)

Glucose oxidase Dough strengtheningA. Monfort, A. Blasco, P. Sanz, J.A. Prieto, (1991)

Lipoxygenase Dough strengthening, bread whitening

A. Monfort, A. Blasco, P. Sanz, J.A. Prieto, (1991)

Protease Biscuits, cookiesT. Godfrey, S.I. West1996

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by Shirley Corriher,2016

Enzyme technology achieves texture modification, enhanced freshness and extended product shelf life, thus earning manufacturing and distribution efficiencies

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Use of fungal/plant enzymes in animal feed

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Industry Enzyme class Application References

Animal feed Phytase Phytate digestibility – phosphorus release

A.K. Kies, K.H.F. van Hemert, W.C. Sauer,2001

Xylanase Digestibility T. Godfrey, S.I. West,1996

β-Glucanase Digestibility

Bedford and Classen 1992, Chesson 1987, Galante et al. 1998b and Walsh et al. 1993

Cellulases, hemicellulases and pectinases

Production and preservation of high quality fodder for ruminants; improving the quality of grass silage; production of transgenic animals

Ali et al. 1995, Hall et al. 1993 and Selmer-Olsen et al. 1993

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Dr. Piyanun Harnpicharnchai et al. 2010

Yeast with phytase on cell surface offers great benefit as feed supplements

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Use of fungal/plant enzymes in beverages

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Industry Enzyme class Application References

Beverage Pectinase De-pectinization, mashing

Galante et al. 1998b, Godfrey and West 1996b and Uhlig 1998

Amylase Juice treatment, low calorie beer

H. Bisgaard-Frantzen et al.1999

β-Glucanase Mashing

Canales et al. 1988, Galante et al. 1998b, Oksanen et al

Acetolactate decarboxylase Maturation (beer)

RS Pereira et al. 2006

LaccaseClarification (juice), flavor (beer), cork stopper treatment,wine

Xu-Feng, 2005

Pectin esterase Improvement in the clarification of cider

Uhlig, 1998

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 All ethanol contained in alcoholic beverages is produced by means of fermentation induced by yeast. So today when anyone kick back with a cold beer or glass of wine, give thanks for some Fantastic Fungi!

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Role of fungal /plant enzyme in making detergents

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Industry Enzyme class Application References

Detergent (laundry and dish wash) Protease Protein stain removal

T. Godfrey, S.I. West,1996

Amylase Starch stain removalM.B. Tobin, C. Gustafsson, G.W. Huisman,2000

Lipase Lipid stain removalT. Godfrey, S.I. West,1996

Cellulasepreferably acid and endoglucanase rich

Cleaning, color clarification, anti-redeposition of non-denim fabrics (cotton)

Galante et al. 1998a, Godfrey 1996, Kumar et al. 1994 and Kumar et al. 1996

Mannanase Mannanan stain removal (reappearing stains)

Fuglsang et al., 1995

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Protease as detergent

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Use of fungal /plant enzymes in pharmaceutical industry

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Industry Enzyme class Application References

Pharmaceutical and clinical amylase Cold swelling,digestive aids , L. A. Underkofler et al,1958

protease Inflamatory enzyme Advances in Applied Microbiology, Volume 6

lipase

streptokinase Wound debridment L. A. Underkofler, et al. 1958

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Enzymes used in textile processing - photo from Novozymes

Because of the properties of enzymes, they make the textile manufacturing process much more environmentally benign. Generally, they:

operate under milder conditions (temperature and pH) than conventional process chemicals – this results in lower energy costs ( up to 120 kg CO2 savings per ton of textile produced);

save water – reduction of water usage up to 19,000 liters per ton of textiles bleached;

are an alternative for toxic chemicals, making wastewater easier and cheaper to treat. National Institute of Environmental Health Sciences2011

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Use of fungal/plant enzymes in pulp and paper industry

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Industry Enzyme class Application References

Pulp and paper Lipase Pitch control, contaminant control

White, J. S. and White, D. C., eds (1997)

Protease Biofilm removalA Sumantha, C Larroche,2006

AmylaseStarch-coating, de-inking, drainage improvement

Rahkamo et al. 1996

Xylanase Bleach boostingM. L. T. M. Polizeli, 2006,

CellulaseDe-inking, drainage improvement, fiber modification.

Rahkamo et al. 1996

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One of the benefits in industry is to harness fungal metabolism to produce paper pulp from rotted wood.

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Use of fungal /plant enzymes in other organic synthesis

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Industry Enzyme class Application References

Organic synthesis Lipase Resolution of chiral alcohols and amides

K. Clausen,2001

AcylaseSynthesis of semisynthetic penicillin

M Gavrilescu,, Y Chisti, (2005)

NitrilaseSynthesis of enantiopure carboxylic acids

C O'reilly, PD Turner-2003

Leather Protease Unhearing, batingRubin, B. and Dennis, E. A., eds (1997)

Lipase De-picklingRubin, B. and Dennis, E. A., eds (1997)

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The enzymatic process uses proteases and is a more eco-friendly alternative. It can be used to recover hair stripped during this process as well as when integrated with the chemical process can reduce the sulphide and lime used by up to 40% or decrease the liming time by half. Leather created using enzymes has also shown more favorable properties such as strength, surface area, cleaner grain and softness.

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Recent research work done on fungal/plant enzymes in Pakistan• Ishfaq, M. et al., Biochemical and Molecular Studies of various

enzymes activity in fungi. (2016). Molecular Breeding, 7(9): 1-16.

• Clinico-Medical Biochemistry, UAF•  i. Prof. Dr. Khalil-ur-Rehman

ii. Dr. Muhammad Anjum Zia

• Planning and Development Cell Ministry of Science & Technology(MoST) ISLAMABAD Oct. (2013)

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Conclusion• Enzymes are being known to mankind since the ancient human

civilization• Big Thanks to progresses in modern biotechnology, so that enzymes

can be developed today for processes where no one would have anticipated an enzyme to be applicable just a decade ago.• In a world with a promptly increasing folks and approaching

enervation of many natural resources, enzyme technology offers a great potential for many industries to help meet the challenges they will aspect in years to come.

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References

• Ishfaq, M. et al., Biochemical and Molecular Studies of various enzymes activity in fungi. (2016). Molecular Breeding, 7(9): 1-16.• Fungi: Its importance in biotechnology -A review on its past, present

and future prospects,Samrat Chakraborty , Upasana Ghosh Somnath Chakraborty. (2010)• Microbial lipases form versatile tools for biotechnology Karl-Erich Jaeger,

Manfred T Reetz

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• Cellulases and related enzymes in biotechnology M.K. Bhat, August 2000, Food Materials Science Division, Institute of Food Research, Norwich Research Park,Colney,

Norwich, NR4 7UA, UK

• Industrial enzyme applications Ole Kirk , Torben Vedel Borchert, Claus Crone Fuglsang,2000 Research and Development, Novozymes A/S, Krogshoejvej 36, 2880 Bagsvaerd, Denmark,

• Microbial lipases form versatile tools for biotechnology Karl-Erich Jaegera, , Manfred T Reetzb,1998

• Enzymes beyond amylase 2/1/2014 - by Laurie Gorton

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