Application of Response Surface Methodology in Medium Optimization to Improve Lactic Acid Production by Lactobacillus paracasei SRCM201474

Gwangsu Ha, Kim Jin Won, Sua Im, Su-Jin Shin, Hee-Jong Yang, Do-Youn Jeong
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Abstract

The aim of this study was to establish the optimal medium composition for enhancing L(+)-lactic acid (LLA) production using response surface methodology (RSM). Lactobacillus paracasei SRCM201474 was selected as the LLA producer by productivity analysis from nine candidates isolated from kimchi and identified by 16S rRNA gene sequencing. Plackett-Burman design was used to assess the effect of eleven media components on LLA production, including carbon (glucose, sucrose, molasses), nitrogen (yeast extract, peptone, tryptone, beef extract), and mineral (NaCl, K2HPO4, MgSO4, MnSO4) materials. Glucose, sucrose, molasses, and peptone were subsequently chosen as promising media for further optimization studies, and a hybrid design experiment was used to establish their optimal concentrations as glucose 15.48 g/l, sucrose 16.73 g/l, molasses 39.09 g/l, and peptone 34.91 g/l. The coefficient of determination of the equation derived from RSM regression for LLA production was mathematically reliable at 0.9969. At optimum parameters, 33.38 g/l of maximum LLA increased by 193% when compared with MRS broth as unoptimized medium (17.66 g/l). Our statistical model was confirmed by subsequent validation experiments. Increasing the performance of LLA-producing microorganisms and establishing an effective LLA fermentation process can be of particular benefit for bioplastic technologies and industrial applications.
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响应面法在副干酪乳杆菌SRCM201474培养基优化中的应用
本研究的目的是利用响应面法(RSM)确定提高L(+)-乳酸(LLA)产量的最佳培养基组成。从9株泡菜中分离得到副干酪乳杆菌SRCM201474作为LLA产生菌,并通过16S rRNA基因测序进行鉴定。采用Plackett-Burman设计评估了11种培养基成分对LLA生产的影响,包括碳(葡萄糖、蔗糖、糖蜜)、氮(酵母提取物、蛋白胨、色氨酸、牛肉提取物)和矿物(NaCl、K2HPO4、MgSO4、MnSO4)材料。随后选择葡萄糖、蔗糖、糖蜜和蛋白胨作为进一步优化研究的有希望的培养基,并通过混合设计实验确定其最佳浓度为葡萄糖15.48 g/l,蔗糖16.73 g/l,糖蜜39.09 g/l,蛋白胨34.91 g/l。由RSM回归得出的LLA产量方程的决定系数为0.9969,在数学上是可靠的。在优化条件下,最大LLA为33.38 g/l,比未优化培养基MRS肉汤(17.66 g/l)提高了193%。我们的统计模型被后续的验证实验所证实。提高产LLA微生物的性能和建立有效的LLA发酵工艺对生物塑料技术和工业应用具有特别的好处。
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