红曲霉壳聚糖的制备-生物化工专业论文.docxVIP

红曲霉壳聚糖的制备摘要 红曲霉壳聚糖的制备 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M92+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O．3％MgS04·7H20 0．2％NaN03 O．1％KI-12P04 l％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 6C、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 一磨碎一除蛋白质(先用2％HCl、沸水浴、固液比1：6，处理时间 1h，离心洗涤；再用15％Na0H溶液、沸水、固液比1：6，处理1h1 一离心一过滤洗涤(至中性)一脱色处理(10％的双氧水溶液(固液 比1：20)、80C水浴、lh)一酸碱不溶物一干燥一脱乙酰化制备壳 聚糖(碱浓度CNaoH 50％、处理温度1306C、处理时间3h、固液比1 ：4)一离心洗涤过滤一浸提(2％乙酸)一沉淀(用20％NaOH调节 滤液的pH值至9)一沉淀物一干燥一壳聚糖。 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 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％t米浆、O．3％MgS04·7H20、0．2％NaN03、0．1％KH2P04。 相应的工艺条件为：培养基初始pH6．0，培养温度30。C、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各壳聚糖的得率为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各壳聚糖得率6．65％，用红曲黄酒糟制备壳 聚糖得率为0．75％。 关键词：红曲霉黑曲霉壳聚糖发酵 The The preparat i on of Oh i tosan by ferment i ng monascus spp 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 M92+and biotin can increase biomass in evidence．The components of fermenting culture for monascus spp．were determined as follows：4％glucose，2％peptone，O．3％ MgS04‘7H20，0．2％NaN03，0．1％KH2P04，1％biotin．The fermentation conditions for producing chitosan was optimized．the optimal condition of fermentation WaS as follows：temperature 34。C，pH 6．0，amount of medium per flask 100ml／500ml，rotation speed ofshaking 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*crash up--crashing cell and wiping protein(firstly 2％HCl，temperature 1 00(2，ratio of solid an