1株血红蛋白分解菌的分离鉴定及其蛋白酶活力测定

Isolation and Determination of Protease Activity of a Haemoglobin-Degrading Bacteria

为了获得能够有效降解利用屠宰场废弃血液的功能菌株,以日喀则地区屠宰场废弃血液堆积土壤样品为材料,将样品稀释涂布接种在血平板上进行分离,挑取水解圈最大的菌落进行平板划线纯化。对分离菌株进行形态学、生化反应试验、16S rDNA序列鉴定并测定其蛋白酶活性。筛选出1株能够高效降解血红蛋白的菌株命名为NwMCC01910042,该分离菌株为革兰阳性杆菌,V-P(Voges-Proskauer)试验阳性,枸橼酸盐利用、淀粉水解、明胶液化、16S rDNA序列系统进化分析显示NwMCC01910042菌株与Bacillus licheniformis ATCC 14580株的序列相似性为99.79%,与Bacillus licheniformis MSL3076株的序列相似性为99.30%,为地衣芽胞杆菌(Bacillus licheniformis),该菌株的16S rDNA序列已提交至GenBank,准入编号为MN 176417,其蛋白酶活力为188.63 U/mL。利用微生物降解生产氨基酸有机肥的关键是筛选蛋白酶的高产菌株,NwMCC01910042株菌有望作为将废弃血污降解为氨基酸的候选功能菌株。

In order to obtain a functional strain that can effectively decompose discarded blood,this experiment take soil samples in slaughterhouses in Shigatse of Tibet where the discarded blood was accumulated as material.The samples were diluted on blood plates for separation.The colony of largest hydrolysis circle was picked out and purified by scratching method on blood plates.The isolated strain was subjected to morphological,biochemical reaction tests,16S rDNA sequence identification for its protease activity determination.A strain capable of efficiently degrading haemoglobin was named as NwMCC01910042.The isolate was Gram-positive bacillus.with V-P(Voges-Proskauer)test positive,could utilize citrate,hydrolyze starch and liquefy gelatin.The phylogenetic analysis of 16S rDNA sequence indicated that strain NwMCC01910042 showed 99.79%sequence similarity to Bacillus licheniformis ATCC 14580 strain and 99.30%sequence similarity to Bacillus licheniformis MSL3076 strain.The 16S rDNA sequence was committed to GenBank with accession number MN 176417.The protease activity of the strain was 188.63 U/mL.The key of the production of organic amino acids fertilizer using microrganisms is to screen protease high-yielding strain,and strain NwMCC01910042 is hopefully to be a candidate functional strain for degrading discarded blood waste into amino acids.