利用从选定乳制品中分离的酵母分离、生产和优化β -半乳糖苷酶

Folake Titilayo Afolabi, Oluwaseyi Peter Adewale, S. Adeyemo
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摘要

β-半乳糖苷酶是一种将乳糖转化为葡萄糖和半乳糖的酶。它缓解了牛奶副产品(乳清)引起的乳糖不耐受和污染问题。本研究旨在从鲜奶、酸奶和当地生产的奶酪中分离和筛选能够产生β-半乳糖苷酶的酵母菌株。从样品中共分离出115种酵母,其中9种酵母分离株具有产生β-半乳糖苷酶的能力,并选择3种进行进一步分析。采用邻硝基苯基-β-D-吡喃半乳糖苷(ONPG)法筛选β-半乳糖苷酶产生菌。马氏克鲁维酵母SLDY–005菌株产生最高的β-半乳糖苷酶。培养36小时后,在30℃、pH 5.5的温度下,它显示出最大的酶活性(277U/mL)。葡萄糖降低了β-半乳糖苷酶的产量,而酵母提取物和尿素被认为是最佳酶合成的合适氮源。对马氏克鲁维酵母SLDY–005生产的粗β-半乳糖苷酶进行了纯化。透析后部分纯化的酶显示出165.12U/mL的比活性,纯化倍数为6.02,产率为45.91%。纯化酶的最适温度为40℃,pH值为6.0。从当地奶酪中分离出的马氏克鲁维酵母菌株是生产β-半乳糖苷酶的潜在候选菌株,可用于解决乳糖不耐受问题。
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Isolation, production and optimisation of beta-galactosidase by utilizing yeasts isolated from selected dairy products
β-galactosidase is an enzyme that converts lactose into glucose and galactose. It alleviates the issue associated with intolerance to lactose and pollution caused as a result of milk by-products (whey). This study aimed to isolate and select yeast strains that can produce β-galactosidase from fresh milk, yoghurt and locally made cheese. A total of 115 yeasts were isolated from the samples, 9 yeast isolates had the ability to produce β-galactosidase and three (3) were selected for further analysis. β-galactosidase producers were screened using Ortho-nitrophenyl-β-D-galactopyranoside (ONPG) assay. Kluyveromyces marxianus strain SLDY – 005 produced the highest β-galactosidase. It showed maximum enzyme activity (277U/mL) at temperature 30oC, pH 5.5, after an incubation period of 36 h. Glucose had a decrease in β-galactosidase production while yeast extract and urea were considered appropriate nitrogen sources for the best enzyme synthesis. Crude β-galactosidase produced by Kluyveromyces marxianus SLDY – 005 was purified. The partially purified enzyme after dialysis showed a specific activity of 165.12 U/mL and had a purification fold of 6.02 and yield of 45.91 %. The purified enzyme had an optimum temperature of 40oC and a pH 6.0. Kluyveromyces marxianus strain isolated from local cheese is a potential candidate for the production of βgalactosidase and it could be used for combating the problem of lactose intolerance.
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