产几丁质酶高原菌的分离鉴定及酶学性质初步研究  

Isolation,identification and enzymatic characterizations of a chitinase-producing bacterium from plateau

作　　者：权淑静[1,2] 杨文玲[1,2] 雷高 王佰涛 刁文涛[1,2] 梁博 刘德海[1,2] QUAN Shujing;YANG Wenling;LEI Gao;WANG Baitao;DIAO Wentao;LIANG Bo;LIU Dehai(Institute of Biology Co.,Ltd.,Henan Academy of Sciences,Zhengzhou 450008,China;Henan Provincial Key Laboratory ofMicrobial Engineering,Zhengzhou 450008,China;Henan High-tech Industry Co.,Ltd.,Zhengzhou 450008,China)

机构地区：[1]河南省科学院生物研究所有限责任公司,河南郑州450008 [2]河南省微生物工程重点实验室,河南郑州450008 [3]河南省高新技术实业有限公司,河南郑州450008

出　　处：《中国酿造》2023年第11期152-156,共5页China Brewing

基　　金：河南省科学院基金项目(220605056);河南省科技攻关计划项目(222102110101);河南省科学院创新团队项目(20230114)。

摘　　要：该研究通过透明圈法进行菌株初筛和几丁质酶活力测定进行复筛,从西藏日喀则地区土壤中分离筛选高产几丁质酶的菌株,通过形态观察、生理生化试验及16S rDNA序列分析对菌株进行鉴定,并对其产酶酶学性质进行初步研究,同时利用薄层层析(TLC)分析其对几丁质的酶解产物。结果表明,从西藏日喀则地区土壤中分离筛选到一株高产几丁质酶的菌株RGZ2-4-5,经鉴定为Massilia aquatica,其几丁质酶活力为242.36 U/mL。该菌株所产胞外几丁质酶的最适反应温度为50℃,最适反应pH值为5.0,在温度30~40℃、pH 5.0~7.0条件下稳定性较好,金属离子Zn^(2+)、Mn^(2+)、Fe^(2+)、Cr^(2+)、Ag^(+)、Co^(2+)及苯甲基磺酰氟(PMSF)对酶活抑制作用显著(P<0.05),而其他金属离子对酶活无显著的促进或抑制作用(P>0.05)。该酶水解几丁质的最终产物为几丁二糖[(GlcNAc)2]和几丁单糖(GlcNAc)。High chitinase-producing strain was isolated and screened from the soil of Shigatse,Tibet,through preliminary screening of transparent circle method and re-screening of chitinase enzyme activity determination.The strain was identified by morphological observation,physiological and biochemical tests,and 16S rDNA sequence analysis.The enzymatic properties of enzyme produced by the strain were preliminary studied,and its enzymatic hydrolysis products for chitin were analyzed using thin-layer chromatography(TLC).The results showed that the strain RGZ2-4-5 with high chitinase activity was isolated and screened from the soil of Shigatse,Tibet,which was identified as Massilia aquatica.The chitinase activity was 242.36 U/ml.The optimal reaction temperature for extracellular chitinase produced by this strain was 50℃,and the optimal reaction pH was 5.0.The enzyme had good stability under conditions of temperature 30-40℃and pH 5.0-7.0.Metal ions Zn^(2+),Mn^(2+),Fe^(2+),Cr^(2+),Ag^(+),Co^(2+)and phenylmethylsulfonyl fluoride(PMSF)had significant inhibitory effects on the enzyme activity(P<0.05),while other metal ions had no significant promoting or inhibitory effects(P>0.05).The final products of chitin hydrolysis by this enzyme were chitobiose[(GlcNAc)2]and chitin monosaccharide(GlcNAc).

关 键 词：几丁质酶 筛选 鉴定 酶学性质 薄层层析 

分 类 号：TS201.3[轻工技术与工程—食品科学] Q939.97[轻工技术与工程—食品科学与工程]