Barman Nirmal Chandra, Zohora Fatema Tuj, Das Keshob Chandra, Mowla Md Golam, Banu Nilufa Akhter, Salimullah Md, Hashem Abu
Department of Biotechnology and Genetic Engineering, Faculty of Applied Science and Technology, Islamic University, Kushtia, 7003, Bangladesh.
Microbial Biotechnology Division, National Institute of Biotechnology (NIB), Ganakbari, Savar, Dhaka, 1349, Bangladesh.
AMB Express. 2017 Sep 21;7(1):181. doi: 10.1186/s13568-017-0462-6.
The study was conducted to select the best promising keratinolytic bacterial strain. A good keratinase positive bacterium isolated from the soil samples of Hazaribagh tannery industrial zone, Dhaka was identified as Arthrobacter genus depending on the conventional techniques and confirmed as Arthrobacter sp. by sequencing 16S rRNA gene. The medium components and culture conditions were optimized to enhance keratinase production through shake flask culture. Keratin and feather powder (10 g/l or 1%) were good substrates for the highest keratinase production along with yeast extract (0.2 g/l or 0.02%) as an organic nitrogen source and potassium nitrate (1 g or 0.1%) as an inorganic nitrogen source. Maximum yield of keratinase was found after 24 h of incubation at 37 °C with an initial pH of 7.0 and inoculums volume 5% under 150 rpm when keratin, yeast extract and potassium nitrate were used as nutrient sources. Keratinase production was more than 5.0-fold increased when all optimized parameters were applied simultaneously. The optimum reaction temperature and pH were determined to be 40 °C and 8.0 respectively for crude keratinase activity. Therefore, Arthrobacter sp. NFH5 might be used for large scale production of keratinase for industrial purposes in less time.
本研究旨在筛选最具潜力的角蛋白分解细菌菌株。从达卡哈扎里巴格制革工业区土壤样本中分离出的一株角蛋白酶阳性细菌,根据传统技术鉴定为节杆菌属,并通过16S rRNA基因测序确认为节杆菌种。通过摇瓶培养优化培养基成分和培养条件,以提高角蛋白酶产量。角蛋白和羽毛粉(10 g/l或1%)是角蛋白酶产量最高的良好底物,同时酵母提取物(0.2 g/l或0.02%)作为有机氮源,硝酸钾(1 g或0.1%)作为无机氮源。当以角蛋白、酵母提取物和硝酸钾作为营养源时,在37℃、初始pH值为7.0、接种量为5%、150 rpm条件下培养24 h后,角蛋白酶产量最高。当同时应用所有优化参数时,角蛋白酶产量提高了5.0倍以上。粗角蛋白酶活性的最佳反应温度和pH值分别确定为40℃和8.0。因此,节杆菌种NFH5可用于在较短时间内大规模工业生产角蛋白酶。