碩士 / 國立臺灣海洋大學 / 食品科學系 / 93 / This study aimed to investigate the best medium for Bacillus subtilis to produce protease, amylase, chitosanase and cellulase. The produced enzymes were selectively collected and employed to hydrolyze mackerel and asparagus (Gracilaria spp.). The antioxidation ability, proliferation of human hybridoma HB4C5 and physiological activation ability on animal cells of the hydrolysates were analyzed. According to the results obtained in this study, the best complex medium for Bacillus subtilis to produce protease, amylase, chitosanase and cellulase was the mixture containing 1% skim milk, 1% soya meal, 0.25% starch, 0.25% K2HPO4, 0.5% NaCl and 0.05% MgCl2. After 3 days incubation in this complex medium at 37oC, the neutral protease, amylase and cellulase activities were 238.6, 166.0 and 1131.1 unit/mL, respectively. During 8 h hydrolysis using the selected enzymes solution, the antioxidant ability of hydrolysate increased gradually and got the highest values after 7 h hydrolysis at 50oC [total antioxidation ability (TEAC): 2.3 mM, α, α-diphenyl-β-picrylhydrazyl (DPPH) scavenging ability: 61.5%, reducing power: 0.75, Fe2+ chelating ability: 75.8%]. However, the antioxidation ability significantly decreased after 8 h hydrolysis (p<0.05). No significant changes in total nitrogen and volatile basic nitrogen (VBN, <25 mg/100 g) was observed during 8 h hydrolysis. Changes in SDS-PAGE and nonproteinous nitrogen suggested that obvious degradation of muscle proteins occurred during hydrolysis. However, the hydrolyzed mackerel could not proliferate the human hybridoma HB4C5 cell and physiologically activate the animal cells. During 8 h hydrolysis of Gracilaria spp. at 50oC, although no increase in DPPH scavenging ability was obtained, the TEAC increased with the duration of hydrolysis (from 0.28 to 0.89mM). According to the changes in reducing sugar, soluble protein and peptides before/after hydrolysis, the selected enzymes from Bacillus subtilis incubated in the mentioned complex medium could effectively hydrolyze the asparagus (Gracilaria spp.).
| Identifer | oai:union.ndltd.org:TW/093NTOU5253008 |
| Date | January 2005 |
| Creators | Pei-Chien Wu, 吳佩蒨 |
| Contributors | , 江善宗, 殷儷容 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 69 |
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