本地土壤纤维素酶产菌分离、筛选及培养基优化

Md. Arju Hossain, Md. Akash Ahammed, Saiful Islam Sobuj, Siratul Kubra Shifat, P. D. Somadder
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引用次数: 4

摘要

纤维素酶在孟加拉国的工业中需求量不断增加。本实验计划在我国发展酶工业,而不是从其他国家进口酶,并从当地土地样品中分离和筛选纤维素降解细菌,并通过培养基优化确保酶的最大产量。在收集了各个Tangail地区的土壤样品后,在羧甲基纤维素板(CMC)上进行了典型的系列稀释,以产生纤维素酶的微生物。根据比例的水解区选择4株分离菌株,分别标记为CBM21、CSG8、CSR35和CHT8,进行形态学和生化分析。然后在250ml的Erlenmeyer烧瓶中使用CMC肉汁在37℃搅拌速率140rpm下发酵48小时,用纤维素酶测定方法对粗酶进行评价。最后,利用不同的发酵工艺参数对分离菌株进行优化,以提高纤维素酶的产量。4个菌株中,CSG8和CBM21的水解值分别为6.96和8.46,清除率最大。这些分离株经鉴定为假单胞菌属(CSG8)。根据筛选过程,以CBM21 (0.156U/ml)、CSG08 (0.106U/ml)和E. coli(阴性对照)为初始酶产率进行优化。优化后,与大肠杆菌(0.44 U/ml)相比,CBM21 (1.35 U/ml)和CSG8 (1.23 U/ml)显著提高。对CBM21和CSG8的有效温度分别为40°C和35°C, pH为7,葡萄糖为5%,孵育时间为24h的菌株进行了表征。之后,这种类型的优化可以成为孟加拉国酶工业的重大举措。
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Cellulase Producing Bacteria Isolation, Screening and Media Optimization from Local Soil Sample
Cellulase enzyme earns consecutively increasing demand in industries of Bangladesh. This experiment has been planned to develop the enzyme industry in our country rather than import enzymes from other countries and conclude with isolating and screening cellulose-degrading bacteria from local land samples and ensuring maximum enzyme production through media optimization. After collecting soil samples from various Tangail areas, a typical dilution of series was performed on a Carboxymethylcellulose plate (CMC) to produce cellulase-producing microorganisms. Based on the hydrolysis zone of the ratio, four isolates were chosen and labeled CBM21, CSG8, CSR35, and CHT8 for further morphological and biochemical analysis. Then subjected to cellulase production in 250 ml of Erlenmeyer flask using CMC broth for 48 hours of fermentation at 37°C at an agitation rate of 140rpm, and cellulase enzyme assay methods evaluated crude enzymes. Eventually, those confirmed isolates were optimized using different parameters by submerged fermentation process for enhanced cellulase production. Among four isolates, CSG8 and CBM21 exhibited the maximum clearance zone with the hydrolytic values 6.96 and 8.46. These isolates were identified as Pseudomonas spp. (CSG8). According to the screening process, CBM21 (0.156U/ml), CSG08 (0.106U/ml), and E. coli (negative control) have been used for optimization with their initial enzyme productivity. After optimization, an amazing enhancement was noticed for CBM21 (1.35 U/ml) and CSG8 (1.23 U/ml) compared to E.coli (0.44 U/ml). The enzyme was characterized in isolates with an effective temperature of 40 and 35°C for CBM21 and CSG8, respectively, and pH 7, glucose 5% with the incubation time 24h was optimal. Aftermath, this type of optimization can be a significant initiative in the enzyme industries of Bangladesh.
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