Download doc211956 - 食品科學系

國立屏東科技大學食品科學系碩士班
研究生專題報告書面摘要
題目：
鹽逆境對納豆菌 2-acetyl-1-pyrroline 合成之影響
指導教授：吳美莉 老師
Effect of salt stress on 2-acetyl-1-pyrroline
報 告 者：簡如玉
formation in Bacillus subtilis ssp. natto
報告日期：97 年 5 月 30 日
摘要
2-acetyl-1-pyrroline (2-AP) 是一種類似爆米花香味的化合物，此化合物為香
米，芋頭、大豆、香蘭與麵包中之重要香氣成分。本實驗目的為探討納豆菌在鹽
逆境下 2-AP 合成機制，故以 RT-PCR 分析 MG 和 △1-pyrroline-5-carboxylicacid
(P5C)合成之基因表現並定量 MG 與 P5C。結果發現，納豆菌於不同鹽濃度誘導
30 分鐘時，甘油-3-磷酸去氫酶 (G3PDH)、三碳糖磷酸異構酶 (TIM)、果糖-1,6二磷酸醛縮酶 (FBA)、MG 合成酶 (MGS)、鳥胺酸轉胺酶 (OAT)、脯胺酸脫氫酶
(PDH) γ-Glutamyl kinase (γ-GK)與 γ-Glutamyl phosphate reductase (γ-GPR)之
mRNA 表現量會隨鹽濃度增加而有明顯遞增的趨勢。另一方面，MG 和 P5C 的
濃度也會隨鹽濃度的增加而增加，且當 1M NaCl 誘導 20 分鐘，MG 蓄積量達最
高(0.44 mmolg-1)而 P5C 蓄積量則為 20.41 μmolg-1。
關鍵字：2-acetyl-1-pyrroline、納豆菌、鹽逆境、methylglyoxal
Abstract
A popcorn-like flavor compound, 2-acetyl-1-pyrroline (2-AP), has been reported
as a potent flavor component of an aromatic rice, taro, pandan and bread. In order to
study the molecular mechanism of 2-AP biosynthesis, we analyzed the expression
profile of MG and P5C biosynthetic enzymes and the level of MG and P5C in
Bacillus subtilis ssp. natto under salt stress. The results showed that the level of
glycerol-3-phosphate dehydrogenase (G3PDH), triosephosphate isomerase (TIM),
fructose-1,6-bisphosphate aldolase (FBA), methylglyoxal synthase (MGS), Ornithne
aminotransferase (OAT), proline dehydrogenase (PDH), γ-Glutamyl kinase (γ-GK)
and γ-Glutamyl phosphate reductase (γ-GPR) mRNA increased significantly after 30
min of exposure at various concentration of NaCl. Otherwise, the concentrations of
MG and P5C increased with increasing NaCl concentration, and when cell were
subjected to 1M NaCl treatment for 20 min, accumulation of MG and P5C were
reached highest (about 0.44 mmolg-1and 20.41 μmolg-1).
Key words: 2-acetyl-1-pyrroline, Bacillus subtilis ssp. natto, salt stress, methylglyoxal
References
1.
廖君綾。2006。酵母在鹽環境下 2-acetyl-1-pyrroline 合成機制探討。國立屏
東科技大學食品科學系碩士論文。
2.
Adams A, Kimpe N D. 2007. Formation of pyrazines and 2-acetyl-1-pyrroline by
Bacillus cereus. Food Chemistry 101:1230-1238.
Aguilera J, Prieto J A. 2001. The Saccharomyces cerevisiae aldose reductase is
implied in the metabolism of methylglyoxal in response to stress conditions.
Current Genetics 39: 273-283.
Aguilera J, Prieto J A. 2004. Yeast cells display a regulatory mechanism in
response to methylglyoxal. FEMS Yeast Research 4:633-641.
Ahmed N, Battah S, Karachalias N, BabaeiJadidi R, Hora´nyi M, Baro´ ti K,
3.
4.
5.
Hollan S, Thornalley P J. 2003. Increased formation of methylglyoxal and
protein glycation, oxidation and nitrosation in triosephosphate isomerase
deficiency. Biochimica et Biophysica Acta 1639: 121-132.
6.
7.
8.
9.
Bradbury L M T, Fitzgerald T L, Henry R J, Jin Q, Waters D L E. 2005. The
gene for fragrance in rice. Plant Biotechnology Journal 3:363-370.
Garay-Arroyo A, Covarrubias A A. 1999. Three Genes Whose Expression is
Induced by Stress in Saccharomyces cerevisiae. Yeast 15:879-892.
Kalapos M P. 1999. Methylglyoxal in living organisms chemistry, biochemistry,
toxicology and biological implications. Toxicology Letters 110 :145-175.
Morita Y, Nakamori S, Takagi H. 2003. L-proline accumulation and freeze
tolerance in Saccharomyces cerevisiae are caused by a mutation in the PRO1
gene encodingγ-glutamyl kinase. Applied and Environmental Microbiology
69:212-219.
10. Pompliano D L, Peyman A, Knowles J R. 1990. Stabilization of a reaction
intermediate as a catalytic device: definition of the functional role of the flexible
loop in triosephosphate isomerase. Biochemistry 29: 3186-3194.
11. Romanczyk LJ, McClelland CA, Post LS, Aitken WM. 1995. Formation of
2-acetyl-1-pyrroline by several Bacillus cereus strains isolated from Cocoa
fermentation boxes. Journal of Agricultural and Food Chemistry 43: 469-75.
12. Thornalley P J. 1996. Pharmacology of methylglyoxal:formation, modification of
proteins and nucleic acids, and enzymatic detoxification a role in pathogenesis
and antiproliferative chemotherapy. General pharmacology 27:565-573.
13. Yoshihashi T, Huong N T T, Inatomi H. 2002. Precursors of 2-acetyl-1-pyrroline,
a potent flavor compound of an aromatic rice variety. Journal of Agricultural and
Food Chemistry 50: 2001-2004.
14. Yoshu Y, Tomohiro K, Kazuo N, Kazuko Y S, Kazuo S. 1997. Regulation of
levels of proline as an osmolyte in plants under water stress. Plant and Cell
Physiology 38:1095-1102.