Enzyme Engineering and Fermentation

As a leading CDMO, BOC Sciences uses enzyme engineering technologies to improve the catalytic activity of enzymes, enhance enzyme tolerance, and enable large-scale production of enzymes. Enzymes produced by microbial fermentation can be used for the enzymatic synthesis of many APIs in the pharmaceutical industry, and some enzymes are also used for therapeutic purposes. We specialize in enzyme engineering and fermentation related projects to meet the needs of our customers in the pharmaceutical industry.

Introduction

Microorganisms-mediated fermentation and biodegradation have been used in the pharmaceutical, food, textile and nutritional industries. Enzymes have been extensively studied as biocatalysts for these biochemical reactions, and the main sources of enzyme are microorganisms. Enzyme engineering is the technology for mass production and application of enzyme preparations. With the development of technologies such as genetic engineering and enzyme isolation and purification, the high cost, complexity and slow reaction time of many enzymes have been improved.

In particular, recombinant DNA technology has greatly improved the production of enzymes compared to natural expression, thus allowing them to be obtained at low cost and in large quantities. In addition, the selection of superior strains, the development of genetically engineered strains and the optimization of fermentation conditions are also important strategies to increase the concentration of enzymes in fermentation broth.

Enzyme Engineering in Pharmaceutical

Enzymes are highly efficient and selective biocatalysts present in living organisms. The unique properties of enzymes make them suitable for many chemical transformations in the pharmaceutical industry, such as group protection and deprotection, selective acylation and deacylation, selective hydrolysis, esterification, ester-exchange, etc. Enzymes have a wide range of promising applications in the production of active pharmaceutical ingredients (APIs) as well as chiral intermediates. In addition, enzymes can be applied to treat human enzyme deficiencies and other diseases such as metabolic disorders, cystic fibrosis, pancreatic insufficiency, and cancer.

Enzyme	Pharmaceutical use
Nitrile hydratase	Nicotinamide
Dehydrogenase	Omapatrilat, tert-leucine, dorzolamide
Amidase, acylase	6-Amino penicillanic acid (6-APA), cefprozil, cefadroxil
Lipase, esterase	Paroxetine, Captopril, posaconazole
Nitrilase	Methylphenidate
Protease	Abacavir
Dehalogenase	Atorvastatin
Adenosine deaminase	Didanosine
Trypsin	Human insulin
D-Hydantoinase	D-Phenylglycine, D-amino acids
D-Amino acid oxidase	Keto acids

Table 1. Examples of the uses of enzyme in pharmaceutical industries¹

Fermentation Production of Enzymes

Most industrial enzymes, including those used in the pharmaceutical industry, are produced by fermentation of suitable microbial strains, including bacteria and fungi, because of their ease of handling, fast growth rate and easy scale-up in the fermenter.

Strain selection: The selection of suitable microbial strains for the production of various enzymes is an important aspect for achieving successful industrial applications. For example, Saccharomyces cerevisiae and Pichia pastoris are microorganisms commonly used for enzyme production. Enzymes are produced by fermentation technology where the enzymes are secreted by microbial strains in the fermentation medium. Enzymes are overexpressed in microbial strains by strain improvement to increase enzyme production.
Fermentation processes: Enzyme production is carried out by two main processes, namely submerged fermentation (SmF) and solid state fermentation (SSF). SmF is carried out in liquid medium, where microorganisms grow and multiply in a suspended state. Typically, SmF is used for the production of extracellular enzyme and overexpressed intracellular enzymes. SSF has several advantages over the SmF, such as high product titers, less wastewater generation, and the ability to use agro-industrial waste as substrate. However, SmF is still the preferred process for enzyme production.

Our Methods

Using high throughput screening (HTS) methods, new biocatalysts can be discovered from microorganisms.
Using recombinant DNA technology, the gene encoding the enzyme is cloned and the target enzyme is heterologously expressed in a microbial strain.
Selection of suitable microbial hosts for the production of various industrial enzymes.
Optimization of fermentation processes to simplify the isolation and purification of enzymes for more economical production of industrial enzymes.

Why Choose BOC Sciences?

Large-scale fermentation capacity of over 100,000 liters
Lowest cost of enzyme production
Highest enzyme availability
Enzyme production process control and tracking
Host strains used are generally recognized as safe (GRAS)

Reference

Chiang, S., Strain improvement for fermentation and biocatalysis processes by genetic engineering technology, J. Ind. Microbiol. Biotechnol., 2004, 31, 99-108.