以杏浆为底物在非无菌培养条件下生产聚半乳糖醛酸酶的研究

Ruhşen Aydın Karaağaç, Mehmet Nuri Aydoğan
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引用次数: 0

摘要

本研究从果园土壤中分离出34株具有果胶降解活性的霉菌。利用这些分离物,获得果胶酶群酶;在批处理系统和非无菌培养条件下进行了果胶裂解酶和聚半乳糖醛酸酶的生产研究。5株聚半乳糖醛酸酶活性菌株编码为:T1、T2、T3、T4、T5,其中T4菌株的聚半乳糖醛酸酶活性最高,为354.4 U/L。因此,使用该分离物继续进行优化分析。以杏浆初始浓度、温度、pH和孵育时间为优化参数。在5 ~ 100 (g/L)范围内改变杏浆初始浓度对酶活性的影响,在50 g/L浓度下测定聚半乳糖醛酸酶活性为397.4 U/L。此外,在pH = 3 ~ 8之间,对每个单元的pH参数进行了分析,并在5 ~ 25℃范围内,通过增加5个单元对温度进行了测试。结果表明,聚半乳糖醛酸酶在pH为5时的最大活性为405.7 U/L,在15°C时的最大活性为406.3 U/L。并在1 ~ 5 d内考察了孵育时间的影响,第4天(96 h)测得聚半乳糖醛酸酶活性最大值为429.0 U/L。因此,进行优化研究的上述T4分离物经分子方法鉴定为Sarocladium strictum (Top ekinküfü) T4。
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Polygalacturonase Production by Sarocladium strictum T4 Isolate using Apricot Pulp as Substrate in Non-Sterile Culture Conditions
In this study, 34 molds showing pectin degradation activity were isolated from the soil of orchards, by several tests. By using these isolates, pectinase group enzymes; studies on pectin lyase and polygalacturonase production were performed in the batch system and under non-sterile culture conditions. 5 isolates denoting polygalacturonase activity were coded as: T1, T2, T3, T4, T5 and the best polygalacturonase activity among these isolates was determined as 354.4 U/L in T4 isolate. Thus, optimization analyses continued by the use of this isolate. Initial apricot pulp concentration, temperature, pH and incubation period were tested as optimization parameters. The effects on enzyme activities were investigated by changing the initial apricot pulp concentration in the range of 5-100 (g/L), and in this regard, polygalacturonase activity was determined as 397.4 U/L at 50 g/L. In addition, the pH parameter was analyzed in each unit between pH = 3 – 8, and the temperature was tested by increasing 5 units in the range of 5-25°C. Consequently, the maximum polygalacturonase activity was determined as 405.7 U/L at pH 5 and 406.3 U/L at 15°C. Besides, the effect of the incubation period was studied within 1-5 days and the maximum polygalacturonase activity was determined as 429.0 U/L on the 4th day (after 96 hours). As a result, the above-mentioned T4 isolate, with which the optimization studies were conducted, was identified as Sarocladium strictum (Top ekinküfü) T4 by molecular methods.
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