以紅麴菌生產葡萄糖胺之培養基組成最佳化研究

Medium Optimization of Producing Glucosamine using Monascus pilosus

魏玉嬌(yu-chiao Wei)

指導教授 ： 吳和生

元智大學/工程學院/化學工程與材料科學學系/碩士(2008年)

https://doi.org/10.6838/YZU.2008.00272

摘要

葡萄糖胺(glucosamine)為一種單胺醣類物質，為人類關節間潤滑結締組織之主要成分，具修復、補充、滋潤、再造受損的軟骨，可治療退化性關節炎。本研究主要是利用微生物醱酵生產葡萄糖胺。討論以Monascus pilosus CCRC-31527為生產紅麴菌株；探討不同碳源、氮源、無機鹽類、酸鹼質及溫度等對菌株生產葡萄糖胺之影響，期望得到紅麴菌生產葡萄糖胺之最佳培養條件，首先進行米糠(rice bran)碳源濃度之探討，尋求米糠複合培養基中三種組成分(米糠25g/L；蔗糖20g/L；氯化胺15g/L)等最佳化濃度探討，以期得到最高產量的葡萄糖胺(glucosamine)。米糠碳源濃度探討之實驗結果顯示，在培養溫度為30℃，轉速200轉，米糠複合培養基之起始酸鹼值為pH 5.0，培養體積為100mL等培養條件下，葡萄糖胺的產量可達到937.5mg/L。

另外，探討醣類為碳源台糖二號砂糖(B-grade white crystal sugar)對菌株Monascus pilosus CCRC-31527生產葡萄糖胺之影響，得到紅麴菌生產葡萄糖胺之最佳培養條件。於搖瓶探討結果顯示，最適培養條件，250 mL三角瓶中培養液100mL，培養基組成份為米糠(25g/L)；台糖二號砂糖(25g/L)；氯化胺(15g/L)，培養基之起始pH值為5.0，在30℃，200 rpm振盪培養第七天，可獲得最佳紅葡萄糖胺為2023.6 mg/L，在以反應曲面法設計及部分因子設計篩選出米糠、台糖二號砂糖、氯化胺為生產培養基之重要成分，可進一步提供小型發酵槽之最佳化培養條件。

關鍵字

紅麴、葡萄糖胺、二級代謝物、碳/氮源、最佳化

並列摘要

Glucosamine is an amino-monosaccha ride and one of the basic
constituents of chitin and chitosan. Glucosamine is the major component of
human inter- articular lubricant connective tissue. It can repair and rebuild
the damaged cartilage and it also can cur e the disease of osteoarthritis. The
objective of this work is to study the parameters affecting the production of
glucosamine hydrochloride by microbial fermentation. We investigated several carbon and nitrogen sources in the medium affecting glucosamine production by Monascus pilosus CCRC 31527 which required a suitable concentration of organic carbon for high glucosamine production. In this study, glucosamine was produced by Monascus pilosus CCRC 31527 in flask culture. The optimum composition for rice powder、sucrose and NH4Cl production was found to be rice bran(25g/L)、sucrose(20g/L) and NH4Cl (15g/L) in the culture of 30℃, pH 5.0 and 100 mL of the complex medium, and 200 rpm in the shaker.. With these compounds, the glucosamine production was 937.5mg/L

It was found that the production of glucosamine using M. pilosus CCRC 31527 was increased by the addition of B-grade white crystal sugar (25g/L), in the culture of 30℃, pH 5.0 and 100 mL of the complex medium, and 200 rpm in the shaker,and was obtained the maximum amount of 2023.6 mg/L. In addition, response surface methodology was used to optimize the concentrations of the rice bran complex medium to evaluate the effects of the composition concentrations on glucosamine productivity. For this kind of complex medium,it would be good for fermentor fermentation of glucosamine production by M. pilosus CCRC 31527.

並列關鍵字

Monascus、Glucosamine、Secondary metabolism、Carbon and nitrate sources、Optimum medium

參考文獻

Performance liquid chromatographic determination of glucosamine in rat plasma. J. Pharm Pharmaceut Sci., 5(2), 176-180(2002).

Anderson, J.W., R.J., Nicolosi, J.F. Borzelleca, Glucosamine effects in humans: a review of effects on glucose metabolism, side effects, safety considerations and efficacy, Food and Chem. Toxicology, 43, 187-201 (2005).

Nakanishi and J. Taira, Dimerumic acid as an antioxidant of the mold: Monascus anka., Free Radical Biology＆Medicine, 28, 999-1004 (2000).

Barclay T.S.,C. Tsourounis and G.M. McCart, Glucosamine.Ann Pharmacother.,32,574-579.(1998)

Baron A.D.,J.S.Zhu and J.S.Zhu,Glucosamine induces insulin resistance in vivo by affecting GLUT4 transolcation in skeletal muscle J. Pharm Pharmaceut Sci., Clin. Invest.,96, 2792-2801 (1995)