分批与流加发酵法生产纤维素酶的研究

采用里氏木霉ＲｕｔＣ－３０，对２．５Ｌ罐分批和流加发酵产酶条件及优化进行了系统研究。通过研究不同浓度的ＳｏｌｋａＦｌｏｃ（纤维素粉）对分批发酵产酶的影响，发现菌体浓度与产酶量随底物浓度增加而增加，当采用５０ｇ／ＬＳｏｌｋａＦｌｏｃ复合１０ｇ／Ｌ麸皮为碳源时，菌体浓度和产酶量最大，最大ＤＣＷ１３．８２ｇ／Ｌ，ＣＭＣａｓｅ２３４．２Ｕ／ｍｌ，ＦＰＡ２１．２５Ｕ／ｍｌ，但ＳｏｌｋａＦｌｏｃ增加至６０ｇ／Ｌ，高浓度底物对菌体初始生长产生强烈抑制，产酶下降。通过研究不同初始Ｓｏｌ－ｋａＦｌｏｃ浓度对流加发酵产酶的影响，发现当初始底物浓度为５０ｇ／ＬＳｏｌｋａＦｌｏｃ复合１０ｇ／Ｌ麸皮时，菌体量和产酶均达到最大值，分别为ＤＣＷ１５．４１ｇ／Ｌ，ＣＭＣａｓｅ３５９．７Ｕ／ｍｌ，ＦＰＡ３０．６Ｕ／ｍｌ，高菌体量是获得纤维素酶高产的关键因素之一。此外用硫铵盐析法对纤维素酶进行了提取。

Cellulase Production by Trichoderma reesei Rut C 30 with Batch and Fed batch Fermentation

Optimization of cellulase production with batch and fed batch culture in 2.5L fermentor by Trichoderma reesei Rut C 30 was carried out. Cellulase production by batch culture with various concentration of Solka Floc were examined. It was found biomass and enzyme titres increased with increasing Solka Floc concentration, 13.82 g/L of maximum DCW, 232.4U/ml of CMCase activities, 21.25U/ml of FPA activities were obtained respectively when 50g/L Solka Floc plus 10g/L wheat bran was used, however when the substrate concentration was further increased to 60g/L Solka Floc plus 10g/L bran, initial cell growth was strong inhibited by high concentration of substrate, result in low enzyme titre. Cellulase production by fed batch culture with various initial concentration of Solka Floc plus 10g/L bran were also investigated, as the best result, maximum biomass, CMCase and FPA of 15.41g/L,359.7U/ml,30.6U/ml were obtained respectively with initial concentration of 50g/L Solka Floc by intermittent fed batch techniques, highcell mass is the one of key factors for high cellulase production. In addition, cellulase was extracted by ammonium sulfate precipitation method.