Abstract: To enhance the yield and the productivity of alkaline poly-galacturonate lyase by Bacillus sp., the kinetic parameters of batch fermentation at different temperatures ranging from 32 to 41℃ were investigated. It was found that the consumption capacity of carbon source increased and the lag stage of cell growth could be shortened with controlling at a higher temperature (41℃). On the other hand, at the lower temperature such as 35℃, the production capability and rate of alkaline poly-galacturonate lyase were promoted. Based on the kinetic parameter analysis, a two-stage temperature controlling strategy, in which temperature was controlled at 41℃ in the first 4 h and then shifted to 35℃, was suggested and experimentally verified. Following this strategy, the following results were obtained by comparison with that of constant temperature 35℃, the maximum alkaline poly-galacturonate lyase activity was 53.0 U/mL, increased by 23.5%, the time of the maximum alkaline poly-galacturonate lyase activity observed was shortened to 16 h, and the productivity was 3315 U/(h×L), enhanced by 39.1%.

Key words: alkaline poly-galacturonate lyase, kinetics, β-amylase, temperature controlling strategy

摘要： 为提高芽孢杆菌WSHB04-02发酵生产碱性果胶酶的产量和生产强度，研究了不同温度(32~41℃)对碱性果胶酶分批发酵动力学参数的影响. 结果表明，较高的温度(41℃)能提高细胞对碳源的利用能力并促进细胞快速进入对数生长期；而较低的温度(35℃)则有利于提高菌体在发酵中后期产酶的能力和速率. 基于不同温度下的动力学参数，提出了分阶段温度控制策略：发酵时间0~4 h时发酵温度为41℃，4 h之后发酵温度为35℃. 经实验验证，采用温度控制策略可使碱性果胶酶的酶活达到53.0 U/mL，生产强度为3315 U/(h·L)，比单一温度模式下的最大值分别提高了23.5%和39.1%.

关键词: 碱性果胶酶, 发酵动力学, 糖化酶, 温度控制策略