Diptera Section. Zoological Survey of India, M-Block, New Alipore, Kolkata, 700053, India.
Department of Microbiology, Bose Institute, P-1/12 CIT Scheme VIIM, Kolkata, 700054, India.
Arch Microbiol. 2021 Aug;203(6):2961-2977. doi: 10.1007/s00203-021-02282-x. Epub 2021 Mar 26.
Structural-genetic characterization of protease producing genes and enzymes from microbial sources are seldom appreciated despite having its substantial utilization in protein engineering or genetic manipulation for biotechnological applications. Aeromonas veronii CMF, a mesophilic bacterium isolated from the gut of Chrysomya megacephala, was found to exhibited significant level of protease activity. For the revelation of genetic potential in relation to protease production, whole genome of this organism was sequenced and analysed while structure-function of different protease enzyme was predicated using various in silico analysis. The 4.5 mb CMF genome was found to encompass various types of protease and mostly they are neutral in nature. Enzyme production was highest in an optimum pH and temperature of 6.0 (32.09 ± 1.015 U/ml) and 35ºC (41.65 ± 1.152 U/ml), respectively. Other culture parameters for optimum production of protease were determined to be inoculum size (1%), incubation period (72 h), shaking condition (125 rpm), carbon and nitrogen source [2% lactose (92.21 ± 3.16 U/ml) and 0.5% urea (163.62 ± 4.31 U/ml), respectively] and effect of surfactants [0.02 mg/ml Tween 80 (174.72 ± 4.48 U/ml)]. Furthermore, A. veronii CMF exhibited significant enzyme production like serine protease (15.22 ± 0.563 U/ml), aspartate protease (33.16 ± 0.762 U/ml) and collagenase (17.26 ± 0.626 U/ml). Genomic information and results of physio-biochemical assays indicate its cost-effective potential use in different enzyme-industry.
尽管在蛋白质工程或遗传操作方面具有重要的应用,但是从微生物来源中对蛋白酶产生基因和酶的结构 - 遗传特征的研究很少受到重视。从大头金蝇肠道中分离到的嗜温细菌气单胞菌 veronii CMF 表现出显著的蛋白酶活性。为了揭示与蛋白酶产生相关的遗传潜力,对该生物体的全基因组进行了测序和分析,同时使用各种计算机分析预测了不同蛋白酶酶的结构 - 功能。发现 4.5 Mb 的 CMF 基因组包含各种类型的蛋白酶,而且大多数都是中性的。在最佳 pH 和温度(分别为 6.0(32.09±1.015 U/ml)和 35°C(41.65±1.152 U/ml))下,酶产量最高。确定蛋白酶最佳生产的其他培养参数为接种量(1%),培养期(72 小时),振荡条件(125 rpm),碳源和氮源[2%乳糖(92.21±3.16 U/ml)和 0.5%尿素(163.62±4.31 U/ml)]和表面活性剂的影响[0.02 mg/ml Tween 80(174.72±4.48 U/ml)]。此外,气单胞菌 veronii CMF 表现出显著的酶产生,如丝氨酸蛋白酶(15.22±0.563 U/ml),天冬氨酸蛋白酶(33.16±0.762 U/ml)和胶原酶(17.26±0.626 U/ml)。基因组信息和生理生化测定结果表明,它具有在不同酶工业中具有成本效益的潜力。