The discovery and use of industrial enzymes in the age of big data.

Big data is ushering in a major transformation of the times, and big data is changing the way we live and understand the world, as well as a source of new inventions and services.
in biotechnology, the rate at which gene sequences can be obtained is unimaginable 10 years ago, thanks to a sharp drop in the cost of sequencing technology.
can we quickly identify and obtain industrially useful target enzyme genes and active enzyme proteins from a large amount of genetic data resources in the shortest possible time? Gene mining (Genomic mining) is based on the need to catalyz specific reactions, from the literature to find the relevant enzyme isogenetic sequence, and as a gene probe, in the genome database for sequence comparing, screening to obtain the coding information of isoenzyme, and then the enzyme batch iso-expression and high-volume screening, and finally obtained a new biocatalyst with better catalytic performance (version 1.0).
With the rapid development of structural biology, people's understanding of protein structure has gradually deepened, coupled with genomics and proteomics to provide a large number of structural and functional information, the molecular transformation of enzymes from the past random mutations, gradually developed to semi-rational or rational design and other more accurate methods, and even the use of computers from the beginning to design enzymes that do not exist in nature.
, the design and preparation of biocatalysts has become easier and simpler, making the requirements for specific enzymes in all industries more urgent and feasible.
this paper will start from today's bio-information big data, different from the traditional methods of industrial enzyme screening new ideas.
, combined with the company's latest research results, a brief introduction to the design of several typical industrial enzymes and mining new strategies.
1. Digging target enzyme genes from sequenced microbial genomes With the rapid development of gene sequencing technology, more and more microbial genomes are sequencing, and the potential function of enzymes encoded by the open reading box of some of these genes has been predicted, but may not be confirmed by experiments;
On the one hand, the annotated hypothentastic enzyme gene can be cloned and expressed directly, and the desired candidate biocatalyst can be obtained through vitality testing, on the other hand, it can be compared and compared with the conservative sequence of the uncommented enzyme, and the potentially functional target neoenzyme coding sequence can be found, and the new target biocatalyst with the structure/function can be obtained by cloning expression.
the latter is relatively high (low success rate), but is more innovative and has easier access to intellectual property.
2. Gene mining diagram 1 based on probe sequence information to excavate new enzymes from the genome database Based on probe sequence information When the relevant enzyme gene (or peptide) sequence that catalyses a certain type of reaction has been documented, this sequence can be used as a gene probe (or template) to search the public genome database to find Candidate enzyme genes with certain ogenesity with the probe sequence, and then design the quotations according to the retrieved gene sequence, obtain the DNA encoding these enzymes by PCR amplification, clone them for expression, and finally, through the target substrate for active screening, i.e. the biocatalyst with specific catalytic functions required may be obtained (Figure 1).
For example, the company selected from the genome database, planned more than a thousand kinds of structure, different functions of alcohol dehydrogenase / niobium-based reductase (Figure 2), has been successfully prepared more than 600 kinds of freeze-dried enzyme preparations, to meet China's pharmaceutical and chemical enterprises for some industrial biocatalyst custom service needs.
2 Molecular evolution tree of alcohol dehydrogenase/carboxyl reductase "thousandase bank" 3. New enzyme gene mining based on the combination of sequence and structural information Directly clones enzyme genes from the sequenced microbial genome and expresses them from heterogeneity, or the technique of digging target enzymes in the gene database based on probe enzyme sequences has matured and achieved good application results, provided that the function of unknown enzymes has been predicted and annotated, or that enzyme gene sequences that catalyz specific substrate transformations have been publicly reported.
However, for certain substrates, enzymes excavated solely on the basis of sequence information of enzymes that catalyz similar reactions often fail to catalytic the transformation of the target substrate, or, although they can catalyz reactions, do not achieve the desired effect.
can be combined with relevant information such as gene mining and structural analysis, it is expected to significantly improve the efficiency of gene mining.
July 2015, The Post published the 2014 Global Industrial Enzyme Industry Research Report and Forecast for the Next Three Years online.
, the global industrial enzyme market has grown year by year, with an annual output growth rate of 5%, reaching US$4.22 billion in 2014.
, the global industrial enzyme market is basically an oligopoly.
in 2014, Novithin, an industrial enzyme giant, had a 44% market share, while DuPont and DSM had 20% and 6%, respectively.
demand varies widely from region to region, with Europe and North America accounting for the largest demand for industrial enzymes, accounting for 80 per cent, while China accounts for only 9.4 per cent.
China's industrial enzyme production reached 1.1657 million tons in 2014 and maintained a 10% annual production growth trend, with production expected to reach 154.87 tons in 2017, 2017, spurred by expanding market demand and good policies.
through the introduction of foreign advanced equipment, excellent strains and the development of new enzyme preparations, China has begun to enter the rapid development of the enzyme preparation industry.
there is still a certain gap in the original innovation of enzyme preparation research and development, and the independent development ability of most enterprises is still very limited.
If we can seize the opportunity of today's era of big data, Internet plus, make full use of the world's shared and shared biological information resources, master the core technology of industrial enzyme design and transformation, and transform it into industrial enzyme real resources with China's independent intellectual property rights, it is expected to lead the rapid rise of China's biodynamical chemistry department from the source, and promote the high starting point and leapfrog development of the bio-manufacturing industry.
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