Zafar Asma, Aftab Muhammad Nauman, Iqbal Irfana, Din Zia Ud, Saleem Mushtaq Ahmad
Faculty of Life Sciences, University of Central Punjab Lahore Pakistan
Institute of Industrial Biotechnology, Government College University Lahore Pakistan
RSC Adv. 2019 Jan 8;9(2):984-992. doi: 10.1039/c8ra06554c. eCollection 2019 Jan 2.
In this study, the industrial applications of a highly thermostable α-amylase as a desizer in the textile industry was evaluated. The cloned gene was expressed in different media (ZBM, LB, ZYBM9, and ZB) with IPTG (isopropyl β-d-1-thiogalactopyranoside) used as an inducer. Lactose was also used as an alternate inducer for the T7 promoter system in . For the large-scale production of the enzyme, different parameters were optimized. The maximum enzyme production was achieved when the volume of medium was 70% of the total volume of fermenter with a 2.0 vvm air supply and 20% dissolved oxygen at a 200 rpm agitation rate. Under all the optimized conditions, the maximum enzyme production was 22.08 U ml min with lactose (200 mM) as an inducer in ZBM medium. The desizing potential of the purified α-amylase enzyme was calculated with different enzyme concentrations (50-300 U ml) at different temperatures (50-100 °C), and pHs (4-9) with varying time intervals (30-120 min). The highest desizing activity was found when 150 U ml enzyme units were utilized at 85 °C and at 6.5 pH for 1 h.
在本研究中,评估了一种高度耐热的α-淀粉酶在纺织工业中作为退浆剂的工业应用。克隆基因在不同培养基(ZBM、LB、ZYBM9和ZB)中表达,使用异丙基-β-D-1-硫代半乳糖苷(IPTG)作为诱导剂。乳糖也用作T7启动子系统的替代诱导剂。为了大规模生产该酶,对不同参数进行了优化。当培养基体积为发酵罐总体积的70%,空气供应量为2.0 vvm,搅拌速度为200 rpm时溶解氧为20%时,实现了最大酶产量。在所有优化条件下,以乳糖(200 mM)作为诱导剂在ZBM培养基中的最大酶产量为22.08 U ml min。在不同温度(50-100°C)、pH值(4-9)和不同时间间隔(30-120分钟)下,用不同酶浓度(50-300 U ml)计算纯化的α-淀粉酶的退浆潜力。当在85°C和pH 6.5下使用150 U ml酶单位1小时时,发现退浆活性最高。