Screening Of Microbial Transglutaminase By New Sterptomyces Mutant

Streptoverticillium mobaraense is widely applied in producing transgultaminase. The cross-links among proteins or peptides are catalysed by transgultaminase derived from a vanity of natural source. Microbial transglutaminase has many advanta ges over other sources-derived transglutaminase resulting in being widely applied in food industry, medicine and biotechnology in the future.1.The strain which could produce transglutamiase,was selected from the soil.In addition,we also tried to construct high transglutaminase-producing E.coli by genetic engineering.In order to identify this strain,the strain patterns,culture characteristics,Physiological and biochemical characteristics were studied,as well as 16SrDNA sequences.The strain had typical morphology of actinomycetes species.The strain has a similar characteristic to Streptomyces mobaraensis in strain patterns,culture characteristics,physiological and biochemical characteristics and 16SrDNA sequences.2.Use the strains of the genome as a template amplification glutaminase gene encoding,using error-prone PCR technology to counter rotating glutaminase encoding genes for random mutation. As wild fungus production to low production, low enzyme activity of glutaminase, substrate specificity is poor, hope that through error-prone PCR counter rotating genes encoding glutaminase,in order to achieve the purpose of improving enzyme activity.3.The transglutaminase gene fragment from Streptomyces mobaraensis was obtained by PCR and error-prone PCR, and then was inserted into mufti-cloning site of pET vector. The recombinant plasmid was then transformed into E.coli.Streptomyces fermentation enzyme production cycle is long, medium and complex, and E.coli is easy to cultivate, grow fast, the advanta ges of clear genetic back ground, will turn glutaminase gene delivery for expression in E.coli, can achieve the purpose of reduce the production cost, shorten the fermentation period.4.Fermentation conditions, such as temperature, speed, time will affect the microbial production ability of glutaminase,so single factor method was used to optimize the culture conditions for E.coli enzyme production, looking for the optimal production conditions,to explore escherichia coli producing glutamine enzyme fermentation production basis.