以 Aspergillus sp. 生產葡萄糖胺之產量最佳化研究

Optimum production of glucosamine using Aspergillus sp.

張玉芬(Yu-Fen Chang)

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.2008.00092

摘要

葡萄糖胺 (glucosamine) 為構成幾丁質 (chitin) 和幾丁聚醣 (chitosan) 之單胺醣類物質。其用途為修復受損的軟骨，並可治療退化性關節炎。本研究主要是利用黃麴菌株 (Aspergillus sp. BCRC 31742)醱酵生產葡萄糖胺。實驗主要分成三部份討論：(1)分析方法，葡萄糖胺分析方式為利用萘異硫氰酸酯(1-naphthyl iso-thiocyanate)為衍生試劑，衍生反應在溫度為50°C時反應1小時，以吡啶為反應溶劑，衍生化合物利用逆向液相層析分離管柱分離，以紫外光檢測器在波長230 nm偵測葡萄糖胺含量，其精密度可達1.00%、準確度可達2.80%；在色層分析之前置方式使用醱酵所得之溼菌塊，以細胞破碎機破碎，經濃度為6 N 之鹽酸在溫度100 °C下反應，於反應4小時後進行葡萄糖胺濃度定量，以同樣前置程序比較溼菌和乾菌的定量準確度，溼菌鹽酸化效果較乾菌佳，反應時間較短；(2)搖瓶培養最佳化培養，探討條件如碳源、氮源、微量元素等，培養基條件以台糖精製細砂( superior white fine granulated sugar )為碳源和複合培養基蛋白質( peptone )為氮源，可得葡萄糖胺濃度為5.48 g/L、生物量為21.6 g/L、含量為0.25 g/g biomass、產率為0.22 g/g carbon source、生產率為32.6 mg/L•h；此外，以複合培養基蛋白質及黃豆為氮源，可得葡萄糖胺濃度為5.41 g/L、生物量為46.67 g/L、含量為0.12 g/g biomass、產率為0.22 g/g carbon source、生產率為32.2 mg/L•h，並將成本降低； (3)以2 L發酵槽進行量產最佳化條件培養，溶氧量設於10%時可得最佳值，迄今可獲得葡萄糖胺濃度為3.91 g/L、生物量為14.6 g/L、含量為0.27 g/g biomass、產率為0.16 g/g carbon source、生產率為23.3 mg/L•h。

關鍵字

葡萄糖胺 ； 黃麴菌屬 ； 鹽酸化 ； 最佳化

並列摘要

Glucosamine (2-amino-2-deoxy-D-glucose) is an amino monosaccharide that is an essential component of mucopolysaccharide and chitin. It can repair and rebuild the damaged cartilage and it also can cure the disease of osteoarthritis. This study used Aspergillus sp. BCRC31742 to product glucosamine by using fermentation. Three parts were discussed in this study. First is the analysis method of glucosamine. The glucosamine was analyzed with 1-naphthyl isothiocyanate (NITC) as derivatizing agent. The reaction was carried out in pyridine at 50oC for 1 h. The derivative was analyzed by means of high performance liquid chromatograph. The analytical precision and the accuracy for glucosamine is below 1.00% and 2.80%, respectively. The proceduce of pretratment was to take 1g cell wet weight into test tubes, add HCl (6N, 10mL3) at 100℃ for 4 hours, and then neutralize the solution with NaOH to pH 7 to obtain glucosamine aqueous solution. Second is the medium optimization in flask culture including kinds of carbon sources, nitrogen sources and macro nutrient was studied. The experimental result shows that the optimum glucosamine concentration of 5.48 g/, biomass of 21.6 g/L, content of 0.25 g/g biomass, yield of 0.22 g/g carbon source and productivity of 32.6 mg/L．h by using Aspergillus sp. BCRC31742 culture in medium, which carbon source is superior white fine granulated sugar and nitrogen source is peptone. On the other hand, the nitrogen sources were soy bean meal and peptone, the glucosamine concentration has a maximum value 5.41 g/L; biomass, 46.7 g/L; content, 0.12 g/g biomass; yield, 0.22g/g carbon source and productivity, 32.2 mg/L．h. Finally, use the flask culture condition to carry out glucosamine fermentation in a fermenter with changing the aeration with air and oxygen to control the dissolved oxygen. When dissolved oxygen was 10%, there are maximum glucosamine concentration 3.91 g/L; biomass conc. 14.6 g/L; content 0.27 g/g biomass; yield 0.16 g/g carbon source; productivity 23.3 mg/L．h.

並列關鍵字

Glucosamine ； Aspergillus sp. ； Hydrochlorination ； Optimum medium

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