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Production of High Concentrations of Fuel Ethanol from Inulin

用菊粉生产高浓度燃料乙醇

基本信息

批准号：
05660102
负责人：
OHTA Kazuyoshi
金额：
$ 1.34万
依托单位：
Miyazaki University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1994
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05660102/
关键词：
Aspergillus niger Saccharomyces cerevisiae Inulin Inulinase Ethanol Jerusalem artichoke 並行複発酵

项目摘要

Two inulinases, P-IA (MW 70,000) and P-IB (MW 68,000), were purified from the culture filtrate of an inulinase-hyperproducing mutant, Aspergillus niger 817, grown on sucrose by DEAE-Cellulofine A-500 and Q-Sepharose HP chromatographies.The specific activities were 350 U/mg for P-IA and 340 U/mg for P-IB.The enzymes were active only toward inulin, and lacked activity toward sucrose, raffinose or levan. The main products of inulin hydrolysis were inulotriose and -tetraose. Thus, both inulinases exhibited the endotype mode of action on inulin. The apparent K_m values for inulin were 0.48 mM for P-IA and 0.50 mM for P-IB at 40ﾟC and pH 5.0.Pure nonhydrolyzed inulin was directly converted to ethanol in a simultaneous saccharification and fermentation process. A.niger 817 was grown in a submerged culture at 30ﾟC for 5 days. The inulin-digestive liquid culture (150 ml) was supplemented with 45 g of inulin, 0.45 g of (NH_4)_2SO_4, and 0.15 g of KH_2PO_4. The medium (pH 5.0) was inoculated with an ethanol-tolerant strain, Saccharomyces cerevisiae 1200, and fermentation was conducted at 30ﾟC.An additional 30 g of inulin was added to the culture after 15 h of fermentation. S.cerevisiae 1200 utilized 99% of the 65 g of inulin during the fermentation, and produced 20.4 and 21.0% (v/v) ethanol from chicory and dahlia inulins, respectively, within 3 days of fermentation. The maximum volumetric productivities of ethanol were 6.2 and 6.0 g/liter/h for chicory and dahlia inulins, respectively. The conversion efficiency of inulin to ethanol was 83 to 84% of the theoretical ethanol yield. Such high concentrations of ethanol were also produced from a inulin-containing agricultural crop, Jerusalem artichoke.

采用DEAE-Cellulofine A-500和Q-Sepharose HP色谱，从菊粉酶高产突变体黑曲霉817的培养滤液中纯化出两种菊粉酶，P-IA （MW 70000）和P-IB （MW 68000）。对P-IA和P-IB的比活性分别为350 U/mg和340 U/mg。这些酶仅对菊粉有活性，对蔗糖、棉子糖和levin缺乏活性。菊糖水解的主要产物为菊糖三糖和-四糖。因此，两种菊粉酶对菊粉均表现出内源性作用模式。在40℃、pH 5.0条件下，P-IA和P-IB的表观K_m值分别为0.48 mM和0.50 mM。在糖化和发酵的同时，将未水解的菊粉直接转化为乙醇。a .niger 817在30℃下培养5天。在150 ml的菊粉消化液培养液中加入45 g菊粉、0.45 g (NH_4)_2SO_4和0.15 g KH_2PO_4。培养基（pH 5.0）接种耐乙醇菌株Saccharomyces cerevisiae 1200，在30℃的温度下发酵。发酵15 h后，在培养液中加入30 g菊粉。酿酒酵母1200在发酵过程中利用了65 g菊粉的99%，菊苣菊粉和大丽花菊粉在发酵3天内分别产生20.4%和21.0% （v/v）的乙醇。菊苣菊和大丽花菊的最大乙醇体积产率分别为6.2和6.0 g/l /h。菊粉制乙醇的转化率为理论乙醇产率的83 ~ 84%。如此高浓度的乙醇也可以从含有菊粉的农作物耶路撒冷洋蓟中生产出来。