Kim Y O, Lee J K, Kim H K, Yu J H, Oh T K
Microbial Enzyme RU, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Taejon, South Korea.
FEMS Microbiol Lett. 1998 May 1;162(1):185-91. doi: 10.1111/j.1574-6968.1998.tb12997.x.
Phytase hydrolyzes phytate to release inorganic phosphate, which would decrease the addition of phosphorus to feedstuffs for monogastric animals and thus reduce environmental pollution. The gene encoding phytase from Bacillus sp. DS11 was cloned in Escherichia coli and its sequence determined. A 560-bp DNA fragment was used as a probe to screen the genomic library. It was obtained through PCR of Bacillus sp. DS11 chromosomal DNA and two oligonucleotide primers based on N-terminal amino acid sequences of the purified protein and the cyanogen bromide-cleaved 21-kDa fragment. The phy cloned was encoded by a 2.2-kb fragment. This gene comprises 1152 nucleotides and encodes a polypeptide of 383 amino acids with a deduced molecular mass of 41,808 Da. Phytase was produced to 20% content of total soluble proteins in E. coli BL21 (DE3) using the pET22b(+) vector with the inducible T7 promoter. This is the first nucleic sequence report on phytase from a bacterial strain.
植酸酶可水解植酸盐以释放无机磷酸盐，这将减少向单胃动物饲料中添加磷的量，从而减少环境污染。克隆了芽孢杆菌属DS11的植酸酶编码基因，并测定了其序列。用一个560bp的DNA片段作为探针筛选基因组文库。它是通过芽孢杆菌属DS11染色体DNA的PCR以及基于纯化蛋白的N端氨基酸序列和溴化氰裂解的21kDa片段设计的两个寡核苷酸引物获得的。克隆的植酸酶基因由一个2.2kb的片段编码。该基因包含1152个核苷酸，编码一个由383个氨基酸组成的多肽，推导分子量为41808Da。使用带有可诱导T7启动子的pET22b(+)载体，在大肠杆菌BL21(DE3)中生产的植酸酶占总可溶性蛋白的20%。这是关于细菌菌株植酸酶的首个核酸序列报道。
