| Summary: | 碩士 === 國立臺灣海洋大學 === 食品科學系 === 103 === Abstract
The aim of this study is to investigate biological activities of defatted soybean flour (DSF) mixed oligosaccharides (Os), okara (O) mixed oligosaccharides, and the lactic acid bacteria fermented DSF oligosaccharides products. Based on the 16S ribosomal RNA sequencing results, bacterial isolates ST-BO-108, ST-BO-109, and ST-BO-224 were identified as Bacillus subtilis. The experimental results indicated that as strains ST-BO-108, ST-BO-109, and ST-BO-224 were induced by DSF or okara, the -amylase activity observed were 29.72, 3.4, and 27.46 U and 23.57, 22.97, and 27.46 U, respectively. 5% (w/v) DSF and 5% (w/v) okara hot-acid extract were hydrolyzed by ST-BO-108, ST-BO-109, and ST-BO-224 crude enzymes individually, then the Os hydrolysates were passed through a UF system to obtain the Os smaller than 3 kDa. The results of thin layer chromatography (TLC) of these < 3 kDa soybean Os indicated that they were composed of sugars of monosaccharide and DP 2-6. The results on antioxidative test of the DSF mixed Os, DSFOs-1083K, DSFOs-1093K, and DSFOs-2243K as well as okara mixed Os, OOs-1083K, OOs-1093K, and OOs-2243K, showed that 10 mg/mL OOs-2243K performed 49.46% DPPH radical scavenging effect (as 69.51 g/mL trolox) and 10 mg/mL DSFOs-1083K and OOs-1083K had ferrous ion chelating abilities equal to 50 g/mL EDTA (90.81%) had. The results on anti-coagulant activity of the six soybean mixed oligosaccharides showed both no inhibition on clotting and no delay on aggregation time of rabbit plasma. In the tests of inhibition effect on disaccharidase activity, OOs-2243K could inhibit 97.55% and 91.23% enzymatic activities while against maltase and sucrose. As to the effect of inhibition on maltase activity, 3 okara mixed oligosaccharides performed better inhibition effects (97%) than 3 DSF mixed oligosaccharides did. The results of angiotensin-converting enzyme (ACE) inhibition test indicated that 2 mg/mL DSFOs-1093K and OOs-1083K showed 51.57% and 26.73% inhibition effect on ACE activity of control group, respectively. The result of the anti- Japanese encephalitis virus Beijing 1 indicated that 30 g/mL DSFOs-1083K and 300 g/mL OOs-1093K could raise host cell viabilities from 34% (control group) to 70% and 49% at co-treatment group. The result of the anti-enterovirus 71 indicated that 300 g/mL DSFOs-1093K and OOs-1083K could raise host cell viability from 46% (control group) to 99% and 83% at post-infection group. In the experimental results of absorption and transportation by Caco-2 cell, DSFOs-1083K and OOs-2243K could raise total sugar concentrations of basolateral fluid from 0.06 mg/mL to 0.68 mg/mL and 0.57 mg/mL after 180 min reaction, and then up to 1.10 mg/mL and 0.66 mg/mL after 270 min reaction, respectively. After 270 min reaction, DSFOs-1093K showed better absorption effect with Caco-2 cell, its total sugar content reached 2.23 mg. The results of DSF mixed oligosaccharides fermented by 28 LAB strains, the result pointed out that strains BCRC10940, 14026, 12323, and 12327 could yield higher titratable acidity than the rest LAB strains used, and the pH value of their fermentation solutions could decrease to below 4.50. After the DSFO was fermented with single or combined of these 4 LAB strains for 5 hr, the pH value of the fermentation solutions decreased to below 4.5, and LAB counts were increased from 6.00 Log CFU/mL to 7.00 Log CFU/mL. As the ACE inhibition activity of DSF mixed oligosaccharides lactic fermented solution (DSFOs-LFS) fermented by single strain or mixed strains (2 or 3) LAB for 5 hr, results showed that 2 mg/mL of DSFOs-LFS-A (BCRC 10940), DSFOs-LFS-B (BCRC 14026), DSFOs-LFS-C (BCRC 12323) and DSFOs-LFS-D (BCRC 12327) had effect for inhibiting 16-42% ACE activity. The result of the anti-enterovirus 71 indicated that 300 g/mL DSFOs-LFS-E could raise host cell viability from 41% (control group) to 85% at post-infection group.
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