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Optimization of xylanase from Pseudomonas mohnii isolated from Simlipal Biosphere Reserve, Odisha, using response surface methodology
Journal of Genetic Engineering and Biotechnology ( IF 3.6 ) Pub Date : 2020-12-11 , DOI: 10.1186/s43141-020-00099-7
Manish Paul , Dipti Pravamayee Nayak , Hrudayanath Thatoi

Xylanase has long been recognized as a widely used industrially important enzyme. There are some bacterial species already reported to produce xylanase which have potent xylanolytic activity towards the use of this enzyme in the production of bioethanol from lignocellulosic biomass. In this view, an efficient xylanolytic bacterial strain was isolated and screened from the soil sample of Simlipal Biosphere Reserve. Enzymatic assay for the xylanase activity was evidenced from the most potent bacterial strain, and the culture condition was optimized for obtaining the maximum enzyme activity. The most potent xylanolytic strain was also identified using biochemical and molecular methods. Nineteen xylanolytic bacteria (SXB1-SXB19) were isolated from Simlipal forest soil samples following dilution plate technique using corn cob xylan-enriched nutrient agar medium and screened for their xylanase-producing ability. Among these isolates, SXB19 showed maximum xylanolytic potential with a halozone size of 2.5 cm as evident in the formation of prominent yellow patches surrounding its growth in xylan-enriched nutrient agar plate. In unoptimized condition, SXB19 showed the highest enzymatic activity of 22.5 IU/ml among the 19 bacterial strains. In order to optimize the culture conditions for maximizing the xylanase production, Box-Behnken design of response surface methodology (RSM) was used. Four variables such as incubation time, pH, substrate (corn cob xylan) concentration, and temperature were considered for the RSM optimization study. From the results, it is evident that in an optimized condition of incubation time 36 h, pH 6.0, xylan concentration 0.5%, and temperature 42.5 °C, the enzyme activity reached a maximum of 152 IU/ml with nearly 6.75 times increase from the unoptimised condition. Besides, xylanase production from SXB19 was considerable in the presence of xylan followed by starch, nitrogen source such as urea followed by yeast extract, and mineral ion sources such as KCl followed by MgSO4 and ZnSO4. From different biochemical tests, 16S rRNA gene sequencing, and phylogenetic analysis, the bacterial strain SXB19 was identified as Pseudomonas mohnii. The isolation of Pseudomonas mohnii, a potent xylanolytic bacterium from Simlipal, is a new report which opens up an opportunity for industrial production of xylanase for bioethanol production and other applications.

中文翻译:

使用响应面法优化从奥里萨邦西姆波利尔生物圈保护区分离的假单胞菌木聚糖酶

木聚糖酶长期以来被认为是广泛使用的工业上重要的酶。已经报道了一些产生木聚糖酶的细菌物种,这些细菌具有强力的木聚糖分解活性,从而对该酶在木质纤维素生物质生产生物乙醇中的应用具有重要意义。从这个角度出发,从Simlipal生物圈保护区的土壤样品中分离并筛选了一种有效的木聚糖分解细菌菌株。从最有效的细菌菌株中证明了木聚糖酶活性的酶促测定,并且优化了培养条件以获得最大的酶活性。还使用生化和分子方法鉴定了最有效的木聚糖分解菌株。按照稀释平板技术,使用富含玉米芯木聚糖的营养琼脂培养基从Simlipal森林土壤样品中分离出19种木聚糖分解细菌(SXB1-SXB19),并筛选其木聚糖酶产生能力。在这些分离物中,SXB19表现出最大的木聚糖分解潜能,卤代酮大小为2.5 cm,这在围绕其富含木聚糖的营养琼脂平板中生长的突出黄色斑块形成时很明显。在未优化的条件下,SXB19在19个细菌菌株中表现出最高的酶活性22.5 IU / ml。为了优化培养条件以最大化木聚糖酶的产量,使用了Box-Behnken设计的响应面方法(RSM)。RSM优化研究考虑了四个变量,例如孵育时间,pH,底物(玉米芯木聚糖)浓度和温度。从结果可以明显看出,在温育时间36小时,pH 6.0,木聚糖浓度0.5%和温度42.5°C的最佳条件下,酶活性最高达到152 IU / ml,比酶促活性提高了近6.75倍。未优化的条件。此外,在存在木聚糖,淀粉,氮源(如尿素,酵母提取物),矿物质离子源(如KCl,硫酸镁和ZnSO4)的情况下,SXB19产生的木聚糖酶的产量很高。通过不同的生化测试,16S rRNA基因测序和系统发育分析,细菌菌株SXB19被鉴定为莫氏假单胞菌。一项新的报告是从Simlipal中分离出一种强力的木聚糖分解细菌Pseudomonas mohnii,这为木聚糖酶的工业化生产提供了机会,可用于生物乙醇生产和其他应用。显然,在温育时间为36 h,pH 6.0,木聚糖浓度为0.5%,温度为42.5°C的最佳条件下,酶活性达到最大152 IU / ml,比未优化条件提高了近6.75倍。此外,在存在木聚糖,淀粉,氮源(如尿素,酵母提取物),矿物质离子源(如KCl,硫酸镁和ZnSO4)的情况下,SXB19产生的木聚糖酶的产量很高。通过不同的生化测试,16S rRNA基因测序和系统发育分析,细菌菌株SXB19被鉴定为莫氏假单胞菌。一项新的报告是从Simlipal中分离出一种强力的木聚糖分解细菌Pseudomonas mohnii,这为木聚糖酶的工业化生产提供了机会,可用于生物乙醇生产和其他应用。显然,在温育时间为36 h,pH 6.0,木聚糖浓度为0.5%,温度为42.5°C的最佳条件下,酶活性达到最大152 IU / ml,比未优化条件提高了近6.75倍。此外,在存在木聚糖,淀粉,氮源(如尿素,酵母提取物),矿物质离子源(如KCl,硫酸镁和ZnSO4)的情况下,SXB19产生的木聚糖酶的产量很高。通过不同的生化测试,16S rRNA基因测序和系统发育分析,细菌菌株SXB19被鉴定为莫氏假单胞菌。一项新的报告是从Simlipal中分离出一种强力的木聚糖分解细菌Pseudomonas mohnii,这为木聚糖酶的工业化生产提供了机会,可用于生物乙醇生产和其他应用。木聚糖浓度为0.5%,温度为42.5°C,酶活性最高达到152 IU / ml,比未优化条件提高了近6.75倍。此外,在存在木聚糖,淀粉,氮源(如尿素,酵母提取物),矿物质离子源(如KCl,硫酸镁和ZnSO4)的情况下,SXB19产生的木聚糖酶的产量很高。通过不同的生化测试,16S rRNA基因测序和系统发育分析,细菌菌株SXB19被鉴定为莫氏假单胞菌。一项新的报告是从Simlipal中分离出一种强力的木聚糖分解细菌Pseudomonas mohnii,这为木聚糖酶的工业化生产提供了机会,可用于生物乙醇生产和其他应用。木聚糖浓度为0.5%,温度为42.5°C,酶活性最高达到152 IU / ml,比未优化条件提高了近6.75倍。此外,在存在木聚糖,淀粉,氮源(如尿素,酵母提取物),矿物质离子源(如KCl,硫酸镁和ZnSO4)的情况下,SXB19产生的木聚糖酶的产量很高。通过不同的生化测试,16S rRNA基因测序和系统发育分析,细菌菌株SXB19被鉴定为莫氏假单胞菌。一项新的报告是从Simlipal中分离出一种强力的木聚糖分解细菌Pseudomonas mohnii,这为木聚糖酶的工业化生产提供了机会,可用于生物乙醇生产和其他应用。从未优化条件增加了75倍。此外,在存在木聚糖,淀粉,氮源(如尿素,酵母提取物),矿物质离子源(如KCl,硫酸镁和ZnSO4)的情况下,SXB19产生的木聚糖酶的产量很高。通过不同的生化测试,16S rRNA基因测序和系统发育分析,细菌菌株SXB19被鉴定为莫氏假单胞菌。一项新的报告是从Simlipal中分离出一种强力的木聚糖分解细菌Pseudomonas mohnii,这为木聚糖酶的工业化生产提供了机会,可用于生物乙醇生产和其他应用。从未优化条件增加了75倍。此外,在存在木聚糖,淀粉,氮源(如尿素,酵母提取物),矿物质离子源(如KCl,硫酸镁和ZnSO4)的情况下,SXB19产生的木聚糖酶的产量很高。通过不同的生化测试,16S rRNA基因测序和系统发育分析,细菌菌株SXB19被鉴定为莫氏假单胞菌。一项新的报告是从Simlipal中分离出一种强力的木聚糖分解细菌Pseudomonas mohnii,这为木聚糖酶的工业化生产提供了机会,可用于生物乙醇生产和其他应用。矿物离子源(如KCl),然后是MgSO4和ZnSO4。通过不同的生化测试,16S rRNA基因测序和系统发育分析,细菌菌株SXB19被鉴定为莫氏假单胞菌。一项新的报告是从Simlipal中分离出一种强力的木聚糖分解细菌Pseudomonas mohnii,这为木聚糖酶的工业化生产提供了机会,可用于生物乙醇生产和其他应用。矿物离子源(如KCl),然后是MgSO4和ZnSO4。通过不同的生化测试,16S rRNA基因测序和系统发育分析,细菌菌株SXB19被鉴定为莫氏假单胞菌。一项新的报告是从Simlipal中分离出一种强力的木聚糖分解细菌Pseudomonas mohnii,这为木聚糖酶的工业化生产提供了机会,可用于生物乙醇生产和其他应用。
更新日期:2020-12-13
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