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Future Prospects of Enzyme Engineering

Future Prospects of Enzyme Engineering

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Published by Megh Raj Bhatt

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Published by: Megh Raj Bhatt on Jan 10, 2010
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CDB,TU
Future prospects of enzyme engineering
1
Future prospects of enzyme engineering
E
nzyme engineering is the recent technology growing rapidly due to its higher application in alot of fields and due to having bright and clear future vision. A most exciting development over the last fewyears is the application of genetic engineering techniques to enzyme technology. There are a number of  properties which may be improved or altered by genetic engineering including the yield and kinetics of theenzyme, the ease of downstream processing and various safety aspects. Enzymes from dangerous or unapproved microorganisms and from slow-growing or limited plant or animal tissue may be cloned intosafe high-production microorganisms. The amount of enzyme produced by a microorganism may beincreased by increasing the number of gene copies that code for it. For example; The engineered cells,aided by the plasmid amplification at around 50 copies per cell, produce penicillin – G – Amidaseconstitutively and in considerably higher quantities than does the fully induced parental strain. Suchincreased yields are economically relevant not just for the increased volumetric productivity but also because of reduced downstream processing costs, the resulting crude enzyme being that much purer. Newenzyme structures may be designed and produced in order to improve on existing enzymes or create newactivities. Much protein engineering has been directed at Subtilisin (from Bacillus amyloliquefaciens), the principal enzyme in the detergent enzyme preparation, Alcalase. This has been aimed at the improvementof its activity in detergents by stabilizing it at even higher temperatures, pH and oxidant strength. A number of possibilities now exist for the construction of artificial enzymes. These are generally synthetic polymersor oligomers with enzyme-like activities, often called synzymes. Enzymes can be immobilized i.e., anenzyme can be linked to an inert support material without loss of activity which facilitates reuse andrecycling of the enzyme.Use of engineered enzyme to form biosensor for the analytical use is also recentactivity among the developed countries. Some enzymes make use in diseases diagnosis so they can begenetically engineered to make the task easier. Thus it is obvious that there is huge scope of the enzymetechnology in the future as well as in present.
Introduction
Enzymes are Organic compounds, produced in the living cells to speed up chemical reaction in the biological systems so that they can take place at relatively lower temperature, but themselves remainapparently unchanged during the process. Therefore enzymes are termed as biocatalysts. Biocatalysts areeither proteins (
enzymes
) or, in a few cases, they may be nucleic acids (
ribozymes
; some RNA moleculescan catalyze the hydrolysis of RNA. Today, we know that enzymes are necessary in all living systems, tocatalyze all chemical reactions required for their survival and reproduction – rapidly, selectively andefficiently. Isolated enzymes can also catalyze these reactions. In the case of enzymes however, thequestion whether they can also act as catalysts outside living systems had been a point of controversyamong biochemists in the beginning of the twentieth century. It was shown at an early stage however thatenzymes could indeed be used as catalysts outside living cells, and several processes in which they wereapplied as biocatalysts have been patented These excellent properties of enzymes are utilized in enzymetechnology. For example, they can be used as biocatalysts to catalyze chemical reactions on an industrialscale in a sustainable manner. Their application covers the production of desired products for all humanmaterial needs (e.g., food, animal feed, pharmaceuticals, fine and bulk chemicals, fibers, hygiene, andenvironmental technology), as well as in a wide range of analytical purposes, especially in diagnostics. Infact, during the past 50 years the rapid increase in our knowledge of enzymes – as well as their biosynthesisand molecular biology – now allows their rational use as biocatalysts in many processes, and in additiontheir modification and optimization for new synthetic schemes and the solution of analytical problemsEnzymes have become big business. They are used in many industrial processes to catalyze biological reactions. Enzymes are exploited in a variety of manufacturing processes such as food processing and for the synthesis of medicines such as antibiotics like artificial penicillin. They are also used
2009/08/27Submitted by: Megh Raj Bhatt
 
CDB,TU
Future prospects of enzyme engineering
2
to clean up factory effluents and pollution in water and soil. Many processes can be made faster andcheaper by using the right enzyme and conditions.Optimum conditions are maintained during factory production by use of bioreactors. These are vesselswhich are designed to provide the ideal environment for reactions involving enzymes or living organisms.Source of enzymes used commercial production is plant, animal and microbial cells. Animal enzymes usedcurrently are lipases, tripsin, rennets etc. Most prevalent plant enzymes are papain, proteases, amylases andsoybean lipoxygenase. These enzymes are used in food industries, for example, papain extracted from papaya fruit is used as meat tenderizer and pancreatic protease in leather softening and manufacture of detergents. In addition microbial enzymes have gained much popularity. Production of primary andsecondary metabolites by microorganism is possible only due to involvement of various enzymes. They areof two types: the extracellular and the intracellular enzymes. There is a wide range of extracellular enzymes produced by pathogenic and saprophytic microorganisms such as cellulose, polymethylegalactouronase, pectinmethylesterase etc. These enzyme helps in establishment in host tissues or decomposition of organicsubstrates. The intracellular enzyme like invertase, uricoxidase, asparaginase are of high economic valueand difficult to extract as they produced inside the cell. They can be extracted by breaking the cells bymeans of a homogenizer or a ball mill and extracted them through the biochemical process.Biotechnology offers an increasing potential for the production of goods to meet various humanneeds. In enzyme technology – a sub-field of biotechnology – new processes have been and are beingdeveloped to manufacture both bulk and high added- value products utilizing enzymes as biocatalysts, inorder to meet needs such as food (e.g., bread, cheese, beer, vinegar), fine chemicals (e.g., amino acids,vitamins), and pharmaceuticals. Enzymes are also used to provide services, as in washing andenvironmental processes, or for analytical and diagnostic purposes. The driving force in the development of enzyme technology, both in academia and industry, has been and will continue to be:The development of new and better products, processes and services to meet these needs; and/or The improvement of processes to produce existing products from new raw materials as biomass.The goal of these approaches is to design innovative products and processes that are not only competitive but also meet criteria of sustainability. A positive effect in all these three fields is required for a sustainable process. Criteria for the quantitative evaluation of the economic and environmental impact are in contrastwith the criteria for the social impact, easy to formulate. In order to be economically and environmentallymore sustainable than an existing processes, a new process must be designed to reduce not only theconsumption of resources (e.g., raw materials, energy, air, water), waste production and environmentalimpact, but also to increase the recycling of waste per kilogram of product.
Sources of enzymes:
 
Biologically active enzymes may be extracted from any living organism. A very wide rangeof sources are used for commercial enzyme production from
 Actinoplanes
to
 Zymomonas
, from spinach to snakevenom. Of the hundred or so enzymes being used industrially, over a half are from fungi and yeast and over a thirdare from bacteria with the remainder divided between animal (8%) and plant (4%) sources. A very much larger number of enzymes find use in chemical analysis and clinical diagnosis. Non-microbial sources provide a larger  proportion of these, at the present time. Microbes are preferred to plants and animals as sources of enzymes because:
1.they are generally cheaper to produce.2.their enzyme contents are more predictable and controllable,3.reliable supplies of raw material of constant composition are more easily arranged, and4.plant and animal tissues contain more potentially harmful materials than microbes, including phenolic compounds (from plants), endogenous enzyme inhibitors and proteases.
Table 1
. Some important industrial enzymes and their sources.EnzymeEC numberSourceIntra/extra-cellular Scale of  productionIndustrial use
2009/08/27Submitted by: Megh Raj Bhatt
 
CDB,TU
Future prospects of enzyme engineering
3
Animal enzymes
Catalase1.11.1.6LiverI -FoodChymotrypsin3.4.21.1PancreasE -LeatheLipase3.1.1.3PancreasE -FoodRennet3.4.23.4AbomasumE+CheeseTrypsin3.4.21.4PancreasE-Leathe
Plant enzymes
Actinidin3.4.22.14Kiwi fruitE-Fooda-Amylase3.2.1.1Malted barleyE+++Brewing b-Amylase3.2.1.2Malted barleyE+++BrewingBromelain3.4.22.4Pineapple latexE -Brewing b-Glucanase 3.2.1.6Malted barleyE ++BrewingFicin3.4.22.3Fig latexE-FoodLipoxygenase1.13.11.12SoybeansI-FoodPapain3.4.22.2Pawpaw latexE++Meat
Bacterial enzymes
a-Amylase3.2.1.1
Bacillus
E+++Starch b-Amylase3.2.1.2
Bacillus
E+StarchAsparaginase3.5.1.1
Escherichia coli
I-HealthGlucose isomerase5.3.1.5
Bacillus
I++Fructose syrupPenicillin amidase3.5.1.11
Bacillus
I-PharmaceuticalProtease3.4.21.14
Bacillus
E+++DetergentPullulanase3.2.1.41
Klebsiella
E-Starch
Fungal enzymes
a-Amylase3.2.1.1
Aspergillus
E++BakingAminoacylase3.5.1.14
Aspergillus
I-PharmaceuticalGlucoamylase3.2.1.3
Aspergillus
E+++StarchCatalase1.11.1.6
Aspergillus
I-FoodCellulase3.2.1.4
Trichoderma
E-Waste
2009/08/27Submitted by: Megh Raj Bhatt

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Cecilia Carpio added this note
I think it is a very interesting contribution, however there is an important reference which is not included: http://www1.lsbu.ac.uk/biology/enztech/ 'Enzyme Technology' written by Martin Chaplin and Christopher Bucke (Cambridge University Press, 1990).
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