Zhao Dan, Wang Yao, Na Jin, Ping Wenxiang, Ge Jingping
a Engineering Research Center of Agricultural Microbiology Technology, Ministry of Education , Heilongjiang University , Harbin , China.
b Key Laboratory of Microbiology, College of Heilongjiang Province, School of Life Sciences , Heilongjiang University , Harbin , China.
Prep Biochem Biotechnol. 2019;49(2):202-207. doi: 10.1080/10826068.2019.1566151. Epub 2019 Feb 8.
Lactic acid bacteria (LAB) is an ideal mannanase source due to the bio-safety guarantee. LAB can heterogeneously express β-mannanase or be directly used as β-mannanase-producing strains. This research originally optimized the fermentation condition for β-mannanase produced by Lactobacillus casei HDS-01. The applicable potential of the crude enzyme in juice clarification was investigated. Two-factorial design screened out three factors, i.e., fermentation time (p = 0.0001), glucose (p = 0.0013), and initial pH (p = 0.0167), which significantly affected L. casei HDS-01 β-mannanase activity. Under the predicted conditions resulting from the central composite design (CCD), i.e., fermentation time 18.23 hr, glucose 12.65 g L, initial pH 5.18, the model reached maximal β-mannanase activity of 81.40 U mL. This model was validated by conducting six repeated experiments and subsequent t-test (p = 0.6308). RSM optimization obtained a 1.33-fold increase in β-mannanase activity. This increase could also be qualitatively detected by larger clearance zone on konjac powder-MRS agar through Congo Red dyeing. The yield and clarity of crude β-mannanase-treated juices from orange, apple, and pear were significantly higher than controls without enzyme treatment. This study conferred a relatively high β-mannanase-producing LAB strain with a high bio-safety level and easy and economical use in juice clarification as well as other food-level fields.
由于生物安全性有保障,乳酸菌(LAB)是一种理想的甘露聚糖酶来源。LAB能够异源表达β-甘露聚糖酶,或直接用作产生β-甘露聚糖酶的菌株。本研究最初优化了干酪乳杆菌HDS-01产生β-甘露聚糖酶的发酵条件。研究了粗酶在果汁澄清中的应用潜力。两因素设计筛选出了三个对干酪乳杆菌HDS-01β-甘露聚糖酶活性有显著影响的因素,即发酵时间(p = 0.0001)、葡萄糖(p = 0.0013)和初始pH值(p = 0.0167)。在中心复合设计(CCD)得出的预测条件下,即发酵时间18.23小时、葡萄糖12.65 g/L、初始pH值5.18时,模型达到了最大β-甘露聚糖酶活性81.40 U/mL。通过进行六次重复实验和随后的t检验(p = 0.6308)对该模型进行了验证。响应面法优化使β-甘露聚糖酶活性提高了1.33倍。通过刚果红染色在魔芋粉-MRS琼脂上形成更大的透明圈也能定性检测到这种提高。用粗β-甘露聚糖酶处理的橙汁、苹果汁和梨汁的产量和澄清度显著高于未经酶处理的对照。本研究赋予了一种具有较高生物安全水平、在果汁澄清以及其他食品级领域使用简便且经济的高产β-甘露聚糖酶LAB菌株。