碩士 / 國立臺灣海洋大學 / 食品科學系 / 97 / The object of this study was clone the Agarase gene AS-IIIb and
PV-Ib into the lactic acid bacteria (LAB) with shuttle vector. And
investigate LAB ferment Monostroma nitidum oligosaccharides (MnOS)
contained milk on the effect of MnOS of their antioxidant
properties, anticoagulant properties, and whitening properties. The results
showed that the effectively of protoplast formation of Ped. pentosaceus
MFS, MFL, Lb. bulgaricus BCRC 10696, and Lb. plantarum BCRC 10069
treated with mutanolysin were 46.3%, 3.4%, 5.6%, and 9.9% respectively.
There was more than 7.3 log CFU/mL LAB kept alive after the protoplast
electroporated by 2.0 kV voltage. The constructions of Agarase AS-IIb and
PV-IB genes and vectors pVA838 and pDG148-Stu by restriction-free (RF)
cloning were not completed. The constructed gene of pDG148-Stu vector
were treated with restricted enzyme Stu, T4 polymerase, and dTTP but the
clone was not observed yet. MnOS was separated by ultrafiltration to
obtain (A) 3-5 kDa fraction contained degree of polymerization (DP) 6, 12,
and 30 with the highest reducing sugar content and sulftate content (0.28
mg/mL and 0.68 mg/mg) and (B) 1-3 kDa fraction contained DP 6. In the
fermented experiment of 0.10%, 0.05%, and 0.02% MnOS contained milks,
Lb. plantarum BCRC 10069 + 12250 group took 16 hr to reached the pH
< 4.6, titratable acidity were 0.67-0.68%, and LAB counts were above 9 log
CFU/mL. Ped. pentosaceus MFL + MFS group took 24 hr to reached the pH
< pH 4.6, titratable acidity were 0.60-0.70%, and LAB counts were above
8.5 log CFU/mL. The MnOS-GAI-0.1% cultured milk had the best
scavenging DPPH free radicals ability 21.73% and MnOS-GBII-0.1% had
the best chelating Fe2+ ability 72.56%. The pH value of two MnOS cultured
milks were fell to 4.47-4.07, and titratable acidity were increased to
0.76%-0.89% during 4oC for 14 days. The abilities of antioxidation were
decreased as storage time increased. The APTT anticoagulants activity test
of rabbit plasma showed the MnOS-GAI-100 �慊/mL group compared with
control which could delay the coagulant time for 20 sec, and decrease the
coagulation effect for 32.4%. The rabbit plasma PT anticoagulants activity test result showed it could decrease the coagulation effect for 2.7%, and on human plasma it decreased 8.7% and 7.5% coagulation effect for APTT and PT. The MnOS cultured milks with better antioxidative ability could
increase the cell viability of NIH-3T3 and CCD-966SK of 124-128% and 122-138%. The inhibition of tyrosinase was increased with increasing the concentration of MnOS cultured milk, and the inhibition were 35-37%. The MnOS cultured milks could inhibited the A375 cell, and the cell viability were 46-87%. The experiment results did not obtain the recombined clone, but the result of protoplast formation and electroporation conditions could be consulted. The MnOS cultured milk had the potential to be developed as
health food product and used as whitening ingredient in cosmetic product.
| Identifer | oai:union.ndltd.org:TW/097NTOU5253076 |
| Date | January 2009 |
| Creators | You-Hao Liao, 廖友豪 |
| 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 | 117 |
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