碩士 / 國立臺灣海洋大學 / 食品科學系 / 101 / The purpose of this study is using Sagassum siliquosum polysaccharide hydrolysates (SPSH) that produced from Sagassum siliquosum dry powder treated with 0.4 N HCl heat-acid extraction, cellulase hydrolysis, amylase hydrolysis, and Sagassum siliquosum polysaccharides induced MA103 and MAEF108 crude enzyme hydrolysis as fermentation substrate for biobutanol production by Clostridium spp. The results of heat-acid extraction with different concentrations of diluted HCl and citric acid indicated that the reducing sugars yield of 0.4 N HCl heat-acid extraction was 12.70%. 15 g Sagassum siliquosum dry powder sequentially treated with 121oC hot water extraction for 20 min with 0.4 N HCl, 13,680 U cellulase hydrolysis, 22,000 U amylase hydrolysis, and 100 U (Alginate lyase) MA103 and MAEF108-Sar crude enzymes hydrolysis, the Sagassum siliquosum polysaccharide hydrolysates (SPSH) content 42.30 mg/mL total sugars and 29.86 mg/mL reducing sugars. Then, distinguished the oligosaccharide (OS) which molecular weight less than 3 kDa by ultrafiltration. The TLC analysis showed that SPSH containing monosaccharides and disaccharides. In HPLC analysis, The monosaccharide content increased gradually with hydrolysis procedures, and the glucose yield of SPSH was 56.77% (g/g dry biomass). In salt tolerance test of Clostridium spp., the growth amount of Clostridium acetobutylicum BCRC10639 and Clostridium beijerinckii BCRC17950 decreased with increasing of salinity. This result indicated that 2 Clostridium spp. grew slowly in medium with salinity > 2%, even can not grow. According to the experimental results of carbohydrate utilization test, C. acetobutylicum BCRC10639 can use arabinose, glucose, and lactose as the main carbon source for fermentation, and C. beijerinckii BCRC17950 can not use arabinose and lactose as the main carbon source. Using 60 g/L glucose fermentation solution as substrate inoculated C. acetobutylicum BCRC10639 to stirring batch fermentation, after 168 hr fermentation the glucose consumption was 41.14 g/L and produced 4.32 g/L acetone, 10.95 g/L butanol, and 1.41 g/L ethanol. The ABE yield of C. acetobutylicum BCRC10639 stirring fermentation in 60 g/L glucose solution were 10%, 27%, and 3% (g/g glucose), respectively. Inoculating 10% C. beijerinckii BCRC17950 into 60 g/L glucose solution for stirring batch fermentation. After 168 hr, the consumption of glucose was 40.99 g/L,and produced 4.11 g/L acetone, 10.38 g/L butanol, and 1.53 g/L ethanol. The ABE yield of C. beijerinckii BCRC17950 were 10%, 25%, and 4% (g/g glucose), respectively. Then, using 3-fold dilution SPSH as substrate for stirring fermentation, inoculated with 10% C. acetobutylicum BCRC10639,after 72 hr the total sugars and reducing sugars decreased form 14.10 and 9.95 mg/mL to 5.61 and 3.72 mg/mL, respectively. The acetone and butanol concentrations in fermentation solution were 0.40 and 0.49 g/L.The yield of acetone and butanol were 0.80% and 0.98% (g/g dry biomass), respectively. The 3-fold dilution SPSH inoculated with 10% C. acetobutylicum BCRC10639 for standing fermentation at 120 hr obtained 0.01% acetone and 0.09% butanol, and the yield of acetone and butanol were 0.06% and 0.46% (g/g dry biomass). Inoculating 10% C. beijerinckii BCRC17950 into 3-fold dilution SPSH for standing fermentation, at 96 hr obtained 0.01% acetone and 0.04% butanol, and the yields were 0.04% and 0.21% (g/g dry biomass), respectively.
| Identifer | oai:union.ndltd.org:TW/101NTOU5253044 |
| Date | January 2013 |
| Creators | Chieh-Chih Chen, 陳建志 |
| Contributors | Chorng-Liang Pan, 潘崇良 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 97 |
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