The industrial production of enzymes

The production of enzymes is a pursuit central to the modern biotechnology industry. Markets for traditional industrial enzymes continue to grow while the continued emphasis on biotechnological endeavours has generated demand for an ever increasing number of additional biocatalysts. The advent of genetic engineering has now facilitated the large-scale production of enzymes and other proteins which are produced naturally only in minute quantities. This development is particularly significant with regard to the production of enzymes and other proteins of therapeutic significance, which are now available in clinically useful quantities.

The level of downstream processing to which any enzyme is subjected is dependent upon its intended application. Industrial enzymes produced in bulk generally require little downstream processing, and hence are relatively crude preparations. Enzymes destined for therapeutic applications are subject to a far higher degree of downstream processing, often incorporating 3–4 chromatographic steps.

While enzymology is one of the longest established branches of the biochemical sciences, it continues to be an area of ongoing, active research. The continual discovery of new enzymes and a greater understanding of previously discovered enzymes and their functional significance suggests many novel applications for these catalytic activities. The intestinal production and utilization of enzymes will continue to be of central importance in the biotechnology industry.     

Advances and challenges in the production of extracellular thermoduric pullulanases by wild-type and recombinant microorganisms: a review

Thermoduric pullulanases, acting as starch-debranching enzymes, are required in many industrial applications, mainly in the production of concentrated glucose, maltose, and fructose syrups. To date, however, a single pullulanase, from Bacillus acidopullulyticus, is available on the market for industrial purposes. This review is an investigation of the major advances as well as the major challenges being faced with regard to optimization of the production of extracellular thermoduric pullulanases either by their original hosts or by recombinant organisms. The critical aspects linked to industrial pullulanase production, which should always be considered, are emphasized, including those parameters influencing solubility, thermostability, and catalytic efficiency of the enzyme. This review provides new insights for improving the production of extracellular thermoduric pullulanases in the hope that such information may facilitate their commercial utilization and potentially be applied to the development of other industrially relevant enzymes.     

Enzyme stabilization by nano/microsized hybrid materials

Immobilization is a key technology for successful realization of enzyme‐based industrial processes, particularly for production of green and sustainable energy or chemicals from biomass‐derived catalytic conversion. Different methods to immobilize enzymes are critically reviewed. In principle, enzymes are immobilized via three major routes (i) binding to a support, (ii) encapsulation or entrapment, or (iii) cross‐linking (carrier free). As a result, immobilizing enzymes on certain supports can enhance storage and operational stability. In addition, recent breakthroughs in nano and hybrid technology have made various materials more affordable hosts for enzyme immobilization. This review discusses different approaches to improve enzyme stability in various materials such as nanoparticles, nanofibers, mesoporous materials, sol–gel silica, and alginate‐based microspheres. The advantages of stabilized enzyme systems are from its simple separation and ease recovery for reuse, while maintaining activity and selectivity. This review also considers the latest studies conducted on different enzymes immobilized on various support materials with immense potential for biosensor, antibiotic production, food industry, biodiesel production, and bioremediation, because stabilized enzyme systems are expected to be environmental friendly, inexpensive, and easy to use for enzyme‐based industrial applications.     

Development of applications of industrial enzymes from Malaysian indigenous microbial sources

Malaysian enzyme industry is considered almost non-existence, although the import volume is large. Realizing the importance of enzymes, encompassing a wide range of applications in bioindustry, the development of home grown technologies for enzyme production and applications becomes one of the national priorities in industrial biotechnology. Enzyme production from indigenous microbial isolates was performed either by submerged or solid state fermentation processes. Based on its wide and unique spectrum of properties, enzymes have been developed for wide applications in various industrial processes. The development of the enzyme catalysed applications is based on the modification of the reaction systems to enhance their catalytic activities. Some of the applications of the industrial enzymes include the fine chemicals production, oleochemicals modification, detergent formulation, enzymatic drinking of waste papers, animal feed formulation and effluent treatment processes. Enzymes have also shown to be successfully used as analytical tool in the determination of compounds in body fluids. Although, most of these enzyme catalysed reactions were performed in aqueous phase, the use of enzymes in organic solvents was found to be significant for the production of new chemicals.     

酶学研究的现状与展望

１分子酶学１．１结构研究截至１９９９年１月在Ｂｒｏｏｋｈａｖｅｎｐｒｏｔｅｉｎｄａｔａｂａｎｋ中已有的８０００多个结构中,有４８００个是酶的结构,其中绝大部分是晶体结构,但用ＮＭＲ方法解出的结构正在迅速增长．多标记和多维技术的发展使得用ＮＭＲ方法解出...     

果胶酶生产及工业应用进展

果胶酶是水解酶家族成员,也是生物技术领域的重要酶,其在全球工业酶市场中所占份额约为25%。果胶酶在工业生产中应用广泛,如植物纤维的脱胶、茶和咖啡的发酵、废水处理、纸浆漂白和动物饲料生产等。在果胶酶的天然来源中,由于微生物具有独特的理化性质,最常被用以生产果胶酶。然而,与许多其他工业酶一样,果胶酶也存在野生菌株产量低、工业生产率低等制约因素,因此,目前果胶酶的研究重点主要集中在如何提高工业规模的生产水平。主要介绍了果胶酶的天然来源,以及在这些来源的基础上通过基因工程改造以获得果胶酶高效表达的最新策略,并概括总结了果胶酶发酵工艺和工业应用,以期为生产具有高活性的果胶酶,提高工业生产的效益奠定理论基础。     

Biocatalysts: application and engineering for industrial purposes

Enzymes are widely applied in various industrial applications and processes, including the food and beverage, animal feed, textile, detergent and medical industries. Enzymes screened from natural origins are often engineered before entering the market place because their native forms do not meet the requirements for industrial application. Protein engineering is concerned with the design and construction of novel enzymes with tailored functional properties, including stability, catalytic activity, reaction product inhibition and substrate specificity. Two broad approaches have been used for enzyme engineering, namely, rational design and directed evolution. The powerful and revolutionary techniques so far developed for protein engineering provide excellent opportunities for the design of industrial enzymes with specific properties and production of high-value products at lower production costs. The present review seeks to highlight the major fields of enzyme application and to provide an updated overview on previous protein engineering studies wherein natural enzymes were modified to meet the operational conditions required for industrial application.     

Biotechnological potential of pectinolytic complexes of fungi

Plant cell wall-degrading enzymes, such as cellulases, hemicellulases and pectinases, have been extensively studied because of their well documented biotechnological potential, mainly in the food industry. In particular, lytic enzymes from filamentous fungi have been the subject of a vast number of studies due both to their advantages as models for enzyme production and their characteristics. The demand for such enzymes is rapidly increasing, as are the efforts to improve their production and to implement their use in several industrial processes, with the goal of making them more efficient and environment-friendly. The present review focuses mainly on pectinolytic enzymes of filamentous fungi, which are responsible for degradation of pectin, one of the major components of the plant cell wall. Also discussed are the past and current strategies for the production of cell wall-degrading enzymes and their present applications in a number of biotechnological areas.     

新型工业酶制剂的进步对生物化学品工业生产过程的影响

工业酶制剂对化工和生物化工产品生产的影响有两方面,一是直接催化生产,另一方面是辅助发酵菌进行生产。以下简单回顾了酶制剂在工业领域的应用现状,注重讨论酶制剂在淀粉糖等方面的直接催化应用和在酒精发酵生产等方面的辅助作用,分析新型酶制剂对生产过程的影响及带来的益处,促进行业的可持续发展。     

Valorization of date palm (Phoenix dactylifera) fruit processing by-products and wastes using bioprocess technology – Review

The date palm Phoenix dactylifera has played an important role in the day-to-day life of the people for the last 7000 years. Today worldwide production, utilization and industrialization of dates are continuously increasing since date fruits have earned great importance in human nutrition owing to their rich content of essential nutrients. Tons of date palm fruit wastes are discarded daily by the date processing industries leading to environmental problems. Wastes such as date pits represent an average of 10% of the date fruits. Thus, there is an urgent need to find suitable applications for this waste. In spite of several studies on date palm cultivation, their utilization and scope for utilizing date fruit in therapeutic applications, very few reviews are available and they are limited to the chemistry and pharmacology of the date fruits and phytochemical composition, nutritional significance and potential health benefits of date fruit consumption. In this context, in the present review the prospects of valorization of these date fruit processing by-products and wastes’ employing fermentation and enzyme processing technologies towards total utilization of this valuable commodity for the production of biofuels, biopolymers, biosurfactants, organic acids, antibiotics, industrial enzymes and other possible industrial chemicals are discussed.     

Biotechnological applications of penicillin acylases: state-of-the-art

This review describes the most recent developments in the biotechnological applications of penicillin acylases. This group of enzymes is involved mainly in the industrial production of 6-aminopenicillanic acid and the synthesis of semisynthetic beta-lactam antibiotics. In addition, penicillin acylases can also be employed in other useful biotransformations, such as peptide synthesis and the resolution of racemic mixtures of chiral compounds. Particular emphasis is placed on advances in detection of new enzyme specificities towards other natural penicillins, enzyme immobilization, and optimization of enzyme-catalyzed hydrolysis and synthesis in the presence of organic solvents.     

Exploration of computational approaches to predict the structural features and recent trends in α-amylase production for industrial applications

Amylases are biologically active enzymes that can hydrolyze starch to produce dextrin, glucose, maltose, and oligosaccharides. The amylases contribute approximately 30% to the global industrial enzyme market. The globally produced amylases are widely used in textile, biofuel, starch processing, food, bioremediation of environmental pollutants, pulp, and paper, clinical, and fermentation industries. The purpose of this review article is to summarize recent trends and aspects of α-amylases, classification, microbial production sources, biosynthesis and production methods, and its broad-spectrum applications for industrial purposes, which will depict the latest trends in α-amylases production. In the present article, we have comprehensively compared the biodiversity of α-amylases in different model organisms ranging from archaea to eukaryotes using in silico structural analysis tools. The detailed comparative analysis: regarding their structure, function, cofactor, signal peptide, and catalytic domain along with their catalytic residues of α-amylases in 16 model organisms were discussed in this paper. The comparative studies on alpha (α) amylases' secondary and tertiary structures, multiple sequence alignment, transmembrane helices, physiochemical properties, and their phylogenetic analysis in model organisms were briefly studied. This review has documented the recent trends and future perspectives of industrially important novel thermophilic α-amylases. In conclusion, this review sheds light on the current understanding and prospects of α-amylase research, highlighting its importance as a versatile enzyme with numerous applications and emphasizing the need for further exploration and innovation in this field.     

Plant-associated endophytic fungi as potential bio-factories for extracellular enzymes: Progress,Challenges and Strain improvement with precision approaches

There is an intricate network of relations between endophytic fungi and their hosts that affects the production of various bioactive compounds. Plant-associated endophytic fungi contain industrially important enzymes and have the potential to fulfil their rapid demand in the international market to boost business in technology. Being safe and metabolically active, they have replaced the usage of toxic and harmful chemicals and hold a credible application in biotransformation, bioremediation and industrial processes. Despite these, there are limited reports on fungal endophytes that can directly cater to the demand and supply of industrially stable enzymes. The underlying reasons include low endogenous production and secretion of enzymes from fungal endophytes which have raised concern for widely accepted applications. Hence, it is imperative to augment the biosynthetic and secretory potential of fungal endophytes. Modern state-of-the-art biotechnological technologies aiming at strain improvement using cell factory engineering as well as precise gene editing like Clustered Regularly Interspaced Palindromic Repeats (CRISPR) and its Associated proteins (Cas) systems which can provide a boost in fungal endophyte enzyme production. Additionally, it is vital to characterize optimum conditions to grow one strain with multiple enzymes (OSME). The present review encompasses various plants-derived endophytic fungal enzymes and their applications in various sectors. Furthermore, we postulate the feasibility of new precision approaches with an aim for strain improvement and enhanced enzyme production.     

Alkaliphiles — from an industrial point of view

Abstract: Organisms with pH optima for growth in excess of pH 9 are defined as alkaliphiles (or sometimes alkalophiles). Alkaliphiles contain prokaryotes, eukaryotes, and archaea. It is clear that many different taxa are represented among the alkaliphiles, and some of them are proposed as new taxa. Alkaliphiles can b isolated from normal environments such as garden soil, although counts of the alkaliphiles are higher in alkaline environments. Alkaliphiles have made a great impact in industrial applications. Biological detergents contain alkaline enzymes, such as alkaline cellulases and/or alkaline proteases that have been produced from alkaliphiles. The current proportion of total world enzyme production destined for the laundry detergents market exceeds 30%. Another important application is the industrial production of cyclodextrin with alkaline cyclomaltodextrin glucanotransferase. This enzyme reduced the production cost and paved the way for its use in large quantities in foodstuffs, chemicals and pharmaceuticals. Besides these applications, there are other possible applications in food and waste treatment industries.     

Recent advances on sources and industrial applications of lipases

Lipases are the industrially important biocatalysts, which are envisioned to have tremendous applications in the manufacture of a wide range of products. Their unique properties such as better stability, selectivity and substrate specificity position them as the most expansively used industrial enzymes. The research on production and applications of lipases is ever growing and there exists a need to have a latest review on the research findings of lipases. The present review aims at giving the latest and broadest overall picture of research and development on lipases by including the current studies and progressions not only in the diverse industrial application fields of lipases, but also with regard to its structure, classification and sources. Also, a special emphasis has been made on the aspects such as process optimization, modeling, and design that are very critical for further scale‐up and industrial implementation. The detailed tabulations provided in each section, which are prepared by the exhaustive review of current literature covering the various aspects of lipase including its production and applications along with example case studies, will serve as the comprehensive source of current advancements in lipase research. This review will be very useful for the researchers from both industry as well as academia in promoting lipolysis as the most promising approaches to intensified, greener and sustainable processes. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 34:5–28, 2018     

Mannanases: microbial sources,production, properties and potential biotechnological applications

Mannans are the major constituents of the hemicellulose fraction in softwoods and show widespread distribution in plant tissues. The major mannan-degrading enzymes are β-mannanases, β-mannosidases and β-glucosidases. In addition to these, other enzymes such as α-galactosidases and acetyl mannan esterases, are required to remove the side chain substituents. The mannanases are known to be produced by a variety of bacteria, fungi, actinomycetes, plants and animals. Microbial mannanases are mainly extracellular and can act in wide range of pH and temperature because of which they have found applications in pulp and paper, pharmaceutical, food, feed, oil and textile industries. This review summarizes the studies on mannanases reported in recent years in terms of important microbial sources, production conditions, enzyme properties, heterologous expression and potential industrial applications.     

Enzymatic lysis of microbial cells

Cell wall lytic enzymes are valuable tools for the biotechnologist, with many applications in medicine, the food industry, and agriculture, and for recovering of intracellular products from yeast or bacteria. The diversity of potential applications has conducted to the development of lytic enzyme systems with specific characteristics, suitable for satisfying the requirements of each particular application. Since the first time the lytic enzyme of excellence, lysozyme, was discovered, many investigations have contributed to the understanding of the action mechanisms and other basic aspects of these interesting enzymes. Today, recombinant production and protein engineering have improved and expanded the area of potential applications. In this review, some of the recent advances in specific enzyme systems for bacteria and yeast cells rupture and other applications are examined. Emphasis is focused in biotechnological aspects of these enzymes.     

Chemical modification in the creation of novel biocatalysts

Enzymes are able to perform a multitude of chemical and biochemical transformations with efficiencies that are typically unrivalled by chemical catalysts. However, these evolved systems may lack breadth or utility in other non-natural applications. Altering enzyme and protein scaffolds through covalent modification can expand the usefulness of native biocatalysts not only for synthetic application but also for therapeutic use. This review summarizes recent developments in the field of chemical modification of enzymes and how they can be applied to synthesis and biological research.     

Engineering Tools for the Development of Recombinant Lactic Acid Bacteria

Lactic acid bacteria (LAB) is mainly used in food fermentation. In addition, LAB fermentation technology has been studied in the development of industrial food additives, nutrients, or enzymes used in food processing. In the field of red biotechnology, LAB is approved and is generally recognized as a safe organism and is considered safe for biotherapeutic treatments. Recent clinical trials have demonstrated the medicinal value of therapeutic recombinant LAB and the suitability of innate mechanisms of secretion and anchoring for therapeutic applications such as antibody or vaccine production. However, the gram‐positive phenotypic trait of LAB creates challenges for genetic modifications when compared to other conventional workhorse bacteria, resulting in exclusive developments of genetic tools for engineering LAB. In this review, several distinct approaches in gene expression for engineering LAB are discussed.     

Enzymatic degradation of cell wall and related plant polysaccharides

Polysaccharides such as starch, cellulose and other glucans, pectins, xylans, mannans, and fructans are present as major structural and storage materials in plants. These constituents may be degraded and modified by endogenous enzymes during plant growth and development. In plant pathogenesis by microorganisms, extracellular enzymes secreted by infected strains play a major role in plant tissue degradation and invasion of the host. Many of these polysaccharide-degrading enzymes are also produced by microorganisms widely used in industrial enzyme production. Most commerical enzyme preparations contain an array of secondary activities in addition to the one or two principal components which have standardized activities. In the processing of unpurified carbohydrate materials such as cereals, fruits, and tubers, these secondary enzyme activities offer major potential for improving process efficiency. Use of more defined combinations of industrial polysaccharases should allow final control of existing enzyme processes and should also lead to the development of novel enzymatic applications.