Study on Submerged Culture of Mushroom Mycelium by Bubble Column Fermentors

气泡柱发酵罐深层培养蘑菇菌丝体的研究

• 批准号: 08555208
• 负责人: KAWAGOE Mikio
• 金额: $ 4.8万
• 依托单位: NARA NATIONAL COLLEGE OF TECHNOLOGY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08555208/
• 关键词: Biochemical Engineering; Bubble Column; Bioreactor; Mushroom; Submerged Culture; Reverse Micelles; Extraction; Separation; 逆ミセル抽出; 膜分離; バイオアクター; バイオセンサー

This study is composed of two parts ; the study on submerged culture of mushroom mycelium by bubble column bioreactors, and the study on separation of protein by reverse micelles.I.Submerged culture of mushroom mycelium by bubble column bioreactors : We devised a novel type of airlift column with a tapered riser which is anticipated to be suitable to the submerged culture of mushroom mycelium. The hydrodynamics (gas holdup and liquid circulation velocity) and the oxygen transfer rate in the airlift column were investigated empirically and theoretically. Conditions to suspend large spherical pellets, which are often formed for the submerged culture of mushroom mycelium, in the standard bubble column were investigated. Next, submerged culture of mushroom mycelium was carried out using two species of mushroom : one is productipn of L-malic acid by Schizophyllum commune, and the other is large-scale culture of Tricholoma matsutake. The optimum conditions of the submerged culture on medium composition, temperature, and air flow rate were found for each mushroom.2. Extraction of protein by reverse micelles : Since mushroom mycelium produces many useful proteins, extraction of protein by reverse micelles was investigated. We found the minimal and optimal AOT concentration for the extraction of lysozyme using reverse micelles. The minimal AOT concentration of a binary protein mixture containing cytocbrome c and lysozyme was found to be smaller than the sum of the minimal AOT concentration of each protein obtained in single protein systems. The addition of low concentration of guanidium salts into the feed solution as a pretreatment decreased the minimal AOT concentration, increased the forward extraction, and enabled the successful recoverry of the protein from the micellar organic phase, while the trehalose addition reduced the forward extraction. Using non-ionic Span 60 reverse micelles, cytochrome c was successfully extracted at concentration higher than the CMC.

本研究由两部分组成;蘑菇菌丝体的鼓泡塔生物反应器的液体培养的研究，和蛋白质的分离的研究通过反胶束。I.蘑菇菌丝体的鼓泡塔生物反应器的液体培养：我们设计了一种新型的气升式塔与锥形提升管，这是预期适合于蘑菇菌丝体的液体培养。对气升塔内的流体力学（气含率和液体循环速度）和氧传递速率进行了实验和理论研究。研究了蘑菇菌丝体液体培养中经常形成的球形大颗粒在标准鼓泡塔中的悬浮条件。其次，利用裂褶菌产L-苹果酸和松茸大规模培养两种蘑菇进行菌丝体深层培养。确定了每种蘑菇液体深层培养的最佳培养基组成、温度和通气量.利用反胶团萃取蛋白质：由于蘑菇菌丝体产生许多有用的蛋白质，因此研究了利用反胶团萃取蛋白质。我们找到了反胶束萃取溶菌酶的最低和最佳AOT浓度。发现含有胞质溴和溶菌酶的二元蛋白质混合物的最小AOT浓度小于在单一蛋白质系统中获得的每种蛋白质的最小AOT浓度的总和。添加低浓度的胍盐作为预处理降低了最小AOT浓度，增加了正向萃取，并使蛋白质从胶束有机相中成功回收，而海藻糖的加入减少了正向萃取。使用非离子型Span 60反胶束，在高于CMC的浓度下成功提取了细胞色素c。

项目成果

M.Imai,T.Natsume,K.Naoe,M.Shimizu,S.Ichikawa and S.Furusaki: "Hydrophilic surroundings requisite for the solubilizatio of proteins related with their hydrophobicity in the AOT reversed micellar extraction" Bioseparation. Vol.6. 325-333 (1997)

M.Imai、T.Natsume、K.Naoe、M.Shimizu、S.Ichikawa 和 S.Furusaki：“AOT 反胶束提取中与疏水性相关的蛋白质溶解所需的亲水环境”生物分离。

O.Ura,K.Naoe,M.Kawagoe and M.Imai: "Extraction conditions for protein in Span 60 reverse micellar system" “Biochemical Engineering:Marching toward the Century of Biotechnology"(Ed. by Z.Y.Shen,F.Ouyang,J.T.Yu,Z.A.Gao),Tsinghua Univ.Press (Beijing). 942-94

O.Ura、K.Naoe、M.Kawagoe 和 M.Imai：“Span 60 反胶束系统中蛋白质的提取条件”“生物化学工程：迈向生物技术的世纪”（Z.Y.Shen、F.Ouyang 编辑， J.T.Yu，Z.A.Gao），清华大学出版社（北京）942-94。

K.Naoe,O.Ura,M.Hattori,M.Kawagoe and M.Imai: "Protein Extraction in reverse micelles of Span 60" In“Advances in Bioseparation Engineerin 1997",(Ed.by The Special Study Group for Bioseparation Engineering of SCEJ),(1998). 91-96 (1998)

K.Naoe、O.Ura、M.Hattori、M.Kawagoe 和 M.Imai：“Span 60 反胶束中的蛋白质提取”，“Advances in Bioseparation Engineeringin 1997”，（由生物分离工程特别研究组编辑） SCEJ），（1998）91-96（1998）。

M.Kawagoe, and T.Maeoka: ""Flow Characteristics of an External-Loop Airlift Column with a Tapered Riser"" Kagaku kogaku Ronbunshu. 22. 629-634 (1996)

M.Kawagoe 和 T.Maeoka：“带有锥形立管的外环空运塔的流动特性”Kagaku kogaku Ronbunshu。

M.Kawagoe, Y.Nakamura, and H.Noda: ""Cultivation of Matsutake Mycelium in Bubble Column Bioreactors"" Chemical Engineering Symposium Series 57 <Horizon of Biochemical Engineering>. 109-112 (1997)

M.Kawagoe、Y.Nakamura 和 H.Noda：““鼓泡塔生物反应器中松茸菌丝体的培养””化学工程研讨会系列 57 <生化工程的视野>。