从摇瓶到中试规模,放线菌 B1 生产 α-半乳糖苷酶的统计优化和顺序放大。

IF 2 4区 生物学 Q3 BIOCHEMICAL RESEARCH METHODS
D John Babu, K Balumahendra, T C Venkateswarulu, T Sathish
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引用次数: 0

摘要

α-半乳糖苷酶(α-GAL)是一类水解酶,可从半乳寡糖和合成底物(如 pNPG)中释放半乳糖。本研究采用统计方法提高了乌坦氏放线菌 B1 在浸没发酵过程中产生 α-GAL 的能力。利用 RSM 的 BBD 对温度、pH 值和接种物百分比对酶分泌的影响进行了优化。利用基于 KLa 的放大策略,将优化后的工艺从摇瓶放大到实验室规模(5 升)和中试规模(30 升)。在温度为 28 °C、pH 值为 6.9、接种量为 6.4% 的条件下,使用 BBD 获得了 62.5 U/mL的最高产量。在实验室规模和中试规模的发酵罐中,成功地进行了放大,产率分别达到 74.4 U/mL和 76.8 U/mL。为验证放大策略,还进行了 TOST 验证,结果显示两个规模的置信度均为 95%,表明放大程序得到了完美执行。通过实施 BBD 和放大策略,总体酶产量显著提高到 76%。这是第一篇探讨从 A. utahensis B1 菌株中放大生产 α-GAL 的文章,为工业应用提供了有价值的见解。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Statistical optimization and sequential scale-up of α-galactosidase production by Actinoplanes utahensis B1 from shake flask to pilot scale.

α-Galactosidase (α-GAL) is a class of hydrolase that releases galactose from galacto-oligosaccharides and synthetic substrates such as pNPG. In this study, the production of α-GAL by Actinoplanes utahensis B1 in submerged fermentation was enhanced by using statistical methods. The effects of temperature, pH, and inoculum percentage on enzyme secretion were optimized using BBD of RSM. The optimized process was scaled up from the shake flask to the laboratory scale (5 L) and to pilot scale (30 L) using KLa based scale-up strategy. By using BBD, a maximum yield of 62.5 U/mL was obtained at a temperature of 28 °C, a pH of 6.9, and an inoculum of 6.4%. Scale-up was performed successfully and achieved a yield of 74.4 U/mL and 76.8 U/mL in laboratory scale and pilot scale fermenters. The TOST was performed to validate the scale-up strategy and the results showed a confidence level of 95% for both scales indicating the perfect execution of scale-up procedure. Through the implementation of BBD and scale-up strategy, the overall enzyme yield has been significantly increased to 76%. This is the first article to explore the scale-up of α-GAL from the A. utahensis B1 strain and provide valuable insights for industrial applications.

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来源期刊
Preparative Biochemistry & Biotechnology
Preparative Biochemistry & Biotechnology 工程技术-生化研究方法
CiteScore
4.90
自引率
3.40%
发文量
98
审稿时长
2 months
期刊介绍: Preparative Biochemistry & Biotechnology is an international forum for rapid dissemination of high quality research results dealing with all aspects of preparative techniques in biochemistry, biotechnology and other life science disciplines.
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