Recombined microorganism glutamine transaminase gene and preparation method thereof

Abstract

The invention relates to a recombined microorganism glutamine transaminase gene of a bacillus and expression thereof, belonging to the field of food industrial biotechnology. After the glutamine transaminase gene from the bacillus is cloned and separated, a lactococcus lactis recombinant expression vector spNZ8048-Tgase containing the gene is established and expressed in a lactococcus lactis N9800 strain; the recombined lactococcus lactis is named L.lactis; after an expression product is separated and purified, a recombined glutamine transaminase with a purity of about 90% is obtained; the activity of the recombined glutamine transaminase reaches 2.2U / mL which is greater than twice the activity of the original strain. Due to the adoption of the lactococcus lactis in production of the glutamine transaminase, the production method of the recombined microorganism glutamine transaminase gene has the advantages of high enzyme-producing activity, low production cost and good safety and is simple to operate, and the expressed enzyme is easy to separate and purify, so that a feasible manner for production of the glutamine transaminase is provided.

Description

technical field [0001] The present invention relates to the field of genetic engineering and protein expression, in particular to the soluble expression of transglutaminase zymogen gene in a lactic acid bacteria expression system, specifically a method for preparing recombinant Bacillus microbial transglutaminase. Background technique [0002] The animal product processing industry is an important industry that connects animal husbandry production with people's living needs. On the one hand, with the rapid development of animal husbandry, my country's animal products show a balance between supply and demand and surplus characteristics, but the processing rate of animal products is only about 5%, which is far lower than 60% to 70% in developed countries. Seriously lagging behind, restricting the rapid and efficient development of animal husbandry. On the other hand, with the increase of people's economic income, the improvement of living standards, the optimization of dietary...

AI Technical Summary

Problems solved by technology

At present, the complete gene encoding TGase and the amino acid sequence of TGase protein have been cloned and sequenced, and the TGase activity level obtained by them is very low, which is far from reaching the level of industrialization; the transglutaminase from microbial sources is not affected by season and season because of its production. Raw material limitation is one of the most likely large-scale production applications in the transglutaminase family. Although there are a variety of microorganisms that can produce transglutaminase in a natural state, most of the enzymes are not high; Breeding by mutation breeding technology is time-consuming and laborious, and it is difficult to screen high-yielding strains of transglutaminase, which results in higher actual production costs and limits the large-scale industrial production of the enzyme

Images