摘要
旨在对苏云金芽胞杆菌几丁质酶基因进行分子鉴定和酶活测定,从产几丁质酶活力高的Bt DLD171菌株中提取基因组DNA,通过PCR法克隆得到几丁质酶chi A73基因(Gen Bank登录号为KJ508093)。结果显示,对chi A73基因进行表达,获得大小约74 k D的表达产物。用荧光底物对表达产物进行酶活测定,发现表达产物只能水解荧光底物4-甲基伞形酮几丁三糖[4-MU-(Glc NAc)3],而不能水解4-甲基伞形酮几丁单糖(4-MU-Glc NAc)。结果表明,Chi A73为一种内切几丁质酶,在p H值为8、温度为40℃时酶活性最高。
In order to identify and detect chitinase activities from Bacillus thuringiensis strains, chitinase gene chiA73 （ GenBank accession number ： KJ508093 ） from genomic DNA of Bt DLD171 producing high-yield chitinase was cloned by PCR. Gene of chitinase chiA73 was expressed and the weight of expression product was approximate 74 kD. The results measured by fluorogenic substrates showed that the expression product of chiA73 hydrolyzed 4-MU- （ GlcNAc ） 3 only, but not 4-MU-GlcNAc. In conclusion, the work indicated that ChiA73 was one of endochitinase, and had the highest activity at pH8 and 40℃.
出处
《生物技术通报》
CAS
CSCD
北大核心
2015年第8期147-152,共6页
Biotechnology Bulletin
基金
转基因生物新品种培育科技重大专项(2014ZX0800913B-002)
国家基础科学人才培养基金资助项目(J1210069)
国家高技术研究发展计划课题(2011AA10A203)
国家自然科学基金项目(31401812)
黑龙江省教育厅科学技术研究项目(12521021)
