【作者】 陈旭峰；

【导师】 周立平；

【作者基本信息】 浙江工业大学 ， 生物化工， 2004， 硕士

【摘要】 本文首先对红曲霉发酵法生产壳聚糖的培养基进行了优化。由碳源、氮源、C／N比与菌体生物量和菌丝体细胞壁得率的单因素试验确定了较优的碳源、氮源以及其比例。并由实验获知在培养基中添加适量的Mg²⁺和生物素，能明显提高菌丝体生物量。经实验红曲霉发酵较佳培养基组分为：4％葡萄糖 2％蛋白胨 0.3％MgSO₄·7H₂O 0.2％NaNO3 0.1％KH₂PO₄ 1％生物素。以此为基准对红曲霉进行发酵，确定其较佳的发酵工艺为：培养温度34℃、培养基初始pH值6.0、培养液装量20％、摇床转速200r／min。 同时经实验探讨了酸碱脱蛋白工艺条件、脱色工艺以及脱乙酰化工艺，确定较优的红曲霉壳聚糖的提取工艺路线为：红曲霉干燥菌体→磨碎→除蛋白质（先用2％HCl、沸水浴、固液比1:6，处理时间1h，离心洗涤；再用15％NaOH溶液、沸水、固液比1:6，处理1h）→离心→过滤洗涤（至中性）→脱色处理（10％的双氧水溶液（固液比1:20）、80℃水浴、1h）→酸碱不溶物→干燥→脱乙酰化制备壳聚糖(碱浓度C_NaOH 50％、处理温度130℃、处理时间3h、固液比1:4)→离心洗涤过滤→浸提（2％乙酸）→沉淀（用20％NaOH调节滤液的pH值至9）→沉淀物→干燥→壳聚糖。 本论文对应用乌衣红曲中的泡盛曲霉发酵生产壳聚糖的工艺也进行了初步研究，结果是黑曲霉较优的发酵培养基组分为:4%葡萄糖、4%玉米浆、0.3%Mgso;·7HZo、0.2%NaNo3、0.1%KHZpo4。相应的工艺条件为:培养基初始pH6.0，培养温度30℃、摇床转速200r/min、培养时间48h。 由红曲霉和黑曲霉废菌体以及黄酒糟进行了提取壳聚糖试验，结果是用提取红曲红后的红曲米废渣制备壳聚糖的得率为2.32%，经红曲红提取后的红曲霉液体发酵菌丝体制备壳聚糖得率5.37%，用黑曲霉生产的柠檬酸废渣所制备壳聚糖得率6.65%，用红曲黄酒糟制备壳聚糖得率为0.75%。更多还原

【Abstract】 The culture medium for producing chitosan by fermenting monascus spp. were optimized. The effects of carbon source、nitrogen source、the their ratio on biomass and mycelia cell wall quantity were studied, so that gained the optimal carbon source、nitrogen source、the their ratio. From the experiments, adding to appropriate Mg2+ and biotin can increase biomass in evidence. The components of fermenting culture for monascus spp. were determined as follows: 4 % glucose, 2%peptone, 0.3% MgSO4 ?7H2O, 0.2%NaNO3,0.1 %KH2PO4, 1 % biotin. The fermentation conditions for producing chitosan was optimized, the optimal condition of fermentation was as follows: temperature 34℃, pH 6.0, amount of medium per flask 100ml/500ml, rotation speed of shaking bed 200rpm.The processing conditions of crashing cell and wiping protein , decolor , taking off acetyl in mycelia cell wall were studied, the optimal extracting processing routine was as follow: thallus→crush up→crashing cell and wiping protein（firstly 2% HC1, temperature 100℃, ratio of solid and liquid 1 : 6, disposing time 1h, centrifugal filtration, secondly 15%NaOH, temperature 100℃, ratio of solid and liquid 1 : 6, disposing time 1h） →centrifugal→filtration and washing to pH 7.0→decolor（10% H2O2 liquor（ratio of solid and liquid 1 : 20）, 80℃, 1h）→not dissolve substance→drying→taking off acetyl(alkali concentration C_NaoH50%, temperature 130℃, ratio of solid and liquid 1 : 4, disposing time 3h) → centrifugal→filtration and washing to pH 7.0→taking up （2%acetic acid） →deposition（20%NaOH adjust to liquor’s pH 9.0）→not dissolve substance→dryness→chitosan.The culture medium for producing chitosan by fermenting Aspergillus niger were optimized. The components of fermenting culture for Aspergillus niger were determined as follows: 4% glucose, 4%corn steep liquor , 0.3% MgSO4 ?7H2O, 0.2%NaNO3 ,0.1%KH2PO4. The optimal condition of fermentation was as follows: temperature 30℃, pH 6.0, rotation speed of shaking bed 200rpm.The experiments on abstracting chitosan from the waste monascus spp., Aspergillus niger and yellow wine mycilium were studied. The yield ratio for abstracting chitosan from hongqu rice that had abstracted hongqu pigments was 2.32%, the yield ratio for abstracting chitosan from monascus spp mycilium that had abstracted hongqu pigments was 5.37%, the yield ratio for abstracting chitosan from waste residue in citric acid fermentation was 6.65 % , and waste residue in yellow wine fermentation was 0.75 %.更多还原