Applied Microbiology Laboratory, School of Life Sciences, Devi Ahilya Vishwavidhaylaya, Indore, Madhya-Pradesh, India.
Lett Appl Microbiol. 2022 Aug;75(2):431-441. doi: 10.1111/lam.13746. Epub 2022 Jun 5.
β-cyclodextrin glucosyltransferase (β-CGTase) is an essential enzyme to catalyse the biotransformation of starch into β-cyclodextrins (β-CD). β-CD has widespread applications in the biomedical, pharmaceutical and food industries. The present study focused on β-CGTase production using an efficient natural microbial strain and statistical production optimization for enhanced production. The isolated organism Bacillus sp. NCIM 5799 was found to be 5 μm short bacilli under FE-SEM and alkalihalophilic in nature. The β-CGTase production was optimized using a combination of Plackett-Burman design (PBD) and Central Composite Design-Response Surface Methodology (CCD-RSM). On PBD screening Na CO , peptone and MgSO .7H O were found to be significant for optimal β-CGTase production, whereas the soluble starch and K HPO concentrations were found to be nonsignificant for β-CGTase production. The significant factors obtained after PBD were further optimized using CCD-RSM design. Peptone was found to have a significant interaction effect with Na CO , and MgSO ·7H O and Na CO exhibited a significant effect on the production of CGTase. The production of β-CGTase was enhanced in the presence of peptone (3%) and Na CO (0·8%). CGTase production obtained was 156·76 U/ml when optimized using CCD-RSM. The final optimized medium (RSM) shows 7·7- and 5·4-fold high productions as compared to un-optimized and one factor at a time production media.
β-环糊精葡萄糖基转移酶(β-CGTase)是一种重要的酶,可催化淀粉向β-环糊精(β-CD)的生物转化。β-CD 在生物医学、制药和食品工业中有广泛的应用。本研究专注于使用高效的天然微生物菌株生产β-CGTase,并进行统计生产优化以提高产量。分离得到的芽孢杆菌 NCIM 5799 在 FE-SEM 下为 5μm 短杆菌,呈碱性嗜盐性。通过 Plackett-Burman 设计(PBD)和中心复合设计-响应面法(CCD-RSM)组合优化β-CGTase 的生产。在 PBD 筛选中,发现 Na2CO3、蛋白胨和 MgSO4·7H2O 对最佳β-CGTase 生产有重要影响,而可溶性淀粉和 K2HPO4 浓度对β-CGTase 生产无显著影响。PBD 后获得的显著因素进一步使用 CCD-RSM 设计进行优化。发现蛋白胨与 Na2CO3 具有显著的相互作用效应,而 MgSO4·7H2O 和 Na2CO3 对 CGTase 的生产有显著影响。在存在蛋白胨(3%)和 Na2CO3(0.8%)的情况下,β-CGTase 的产量得到提高。使用 CCD-RSM 优化后,β-CGTase 的产量达到 156.76U/ml。最终优化的培养基(RSM)比未优化和一次优化一种因素的培养基显示出 7.7-和 5.4 倍的高产量。