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利用添加乳清的替代培养基优化乳球菌 UQ2 生产乳链菌肽。

Optimization of nisin production by Lactococcus lactis UQ2 using supplemented whey as alternative culture medium.

机构信息

DIPA, Facultad de Química, Univ Autónoma de Querétaro, Qro, México.

出版信息

J Food Sci. 2010 Aug 1;75(6):M347-53. doi: 10.1111/j.1750-3841.2010.01670.x.

Abstract

Lactococcus lactis UQ2 is a nisin A-producing native strain. In the present study, the production of nisin by L. lactis UQ2 in a bioreactor using supplemented sweet whey (SW) was optimized by a statistical design of experiments and response surface methodology (RSM). In a 1st approach, a fractional factorial design (FFD) of the order 2(5-1) with 3 central points was used. The effect on nisin production of air flow, SW, soybean peptone (SP), MgSO(4)/MnSO(4) mixture, and Tween 80 was evaluated. From FFD, the most significant factors affecting nisin production were SP (P = 0.011), and SW (P = 0.037). To find optimum conditions, a central composite design (CCD) with 2 central points was used. Three factors were considered, SW (7 to 10 g/L), SP (7 to10 g/L), and small amounts of added nisin as self-inducer (NI 34.4 to 74.4 IU/L). Nisin production was expressed as international units (IU). From RSM, an optimum nisin activity of 180 IU/mL was predicted at 74.4 IU/L NI, 13.8 g/L SP, and 14.9 or 5.11 g/L SW, while confirmatory experiments showed a maximum activity of 178 +/- 5.2 IU/mL, verifying the validity of the model. The 2nd-order model showed a coefficient of determination (R(2)) of 0.828. Optimized conditions were used for constant pH fermentations, where a maximum activity of 575 +/- 17 IU/mL was achieved at pH 6.5 after 12 h. The adsorption-desorption technique was used to partially purify nisin, followed by drying. The resulting powder showed an activity of 102150 IU/g. Practical Application: Nisin production was optimized using supplemented whey as alternative culture medium, using a native L. lactis UQ2 strain. Soybean peptone, SW, and subinhibitory amounts of nisin were successfully employed to optimize nisin production by L. lactis UQ2. Dried semipurified nisin showed an activity of 102150 IU/g.

摘要

乳球菌乳 UQ2 是一种产乳链菌素 A 的原生菌株。在本研究中,通过实验设计和响应面法(RSM)的统计设计,对乳球菌乳 UQ2 在补充甜乳清(SW)的生物反应器中的产乳链菌素 A 进行了优化。在第一种方法中,使用阶数为 2(5-1)的部分因子设计(FFD),并带有 3 个中心点。评估了空气流量、SW、大豆蛋白胨(SP)、MgSO4/MnSO4 混合物和 Tween 80 对乳链菌素 A 生产的影响。从 FFD 中,对乳链菌素 A 生产影响最大的因素是 SP(P = 0.011)和 SW(P = 0.037)。为了找到最佳条件,使用带有 2 个中心点的中心组合设计(CCD)。考虑了三个因素,SW(7 至 10 g/L)、SP(7 至 10 g/L)和少量添加的作为自诱导剂的乳链菌素 A(NI 34.4 至 74.4 IU/L)。乳链菌素 A 生产以国际单位(IU)表示。从 RSM 中,预测在 NI 74.4 IU/L、SP 13.8 g/L 和 SW 14.9 或 5.11 g/L 时,乳链菌素 A 的最佳活性为 180 IU/mL,而验证实验表明最大活性为 178 +/- 5.2 IU/mL,验证了模型的有效性。二阶模型的确定系数(R2)为 0.828。在恒 pH 发酵中使用优化条件,在 12 h 后达到 pH 6.5 时,最大活性为 575 +/- 17 IU/mL。采用吸附-解吸技术对乳链菌素 A 进行部分纯化,然后干燥。所得粉末的活性为 102150 IU/g。实际应用:使用补充乳清作为替代培养基,使用原生乳球菌乳 UQ2 菌株,优化了乳链菌素 A 的生产。成功地使用大豆蛋白胨、SW 和亚抑菌量的乳链菌素 A 来优化乳球菌乳 UQ2 的乳链菌素 A 生产。干燥的半纯化乳链菌素 A 显示出 102150 IU/g 的活性。

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