本文選題：綠原酸 + 蘋果�。� 參考：《中國農(nóng)業(yè)大學》2017年博士論文

【摘要】：酚類物質(zhì)是植物體中重要的次級代謝產(chǎn)物,其在人體中的抗氧化、抗衰老、預防心血管疾病以及改善認知等作用已廣泛獲得研究者的關注,但是有關酚類物質(zhì)對果實采后成熟衰老進程是否以及如何產(chǎn)生影響的研究還比較少。本文主要研究果實內(nèi)源酚類物質(zhì)如何對果實成熟衰老相關蛋白質(zhì)的表達以及相應酶活性產(chǎn)生影響從而影響果實成熟衰老進程。研究內(nèi)容與結果如下:實驗發(fā)現(xiàn)相比于新綠原酸(油桃果實果肉組織中另一種主要酚類物質(zhì)),綠原酸可以更顯著地減少油桃果肉切片乙烯釋放量、延緩切片硬度和可溶性固形物含量下降。綠原酸可以顯著降低油桃果實在貯藏期間的乙烯釋放量、呼吸速率、軟化程度、腐爛率和丙二醛含量。同時,綠原酸處理也延緩了貯藏期間油桃果實果皮顏色的轉紅、可滴定酸含量下降和可溶性固形物上升。研究還發(fā)現(xiàn),綠原酸處理顯著增強了果實的抗氧化活性、自由基清除能力和還原能力。實驗利用雙向電泳以及基質(zhì)輔助激光解析電離飛行二級質(zhì)譜(MALDI-TOF-TOF/MS)技術成功分離鑒定了油桃果實中74個與成熟衰老相關并受到綠原酸影響的蛋白質(zhì)。其中參與果實呼吸作用的蛋白質(zhì)NADP-蘋果酸酶(NADP-ME)、蘋果酸脫氫酶和NAD-依賴型蘋果酸脫氫酶,其蛋白表達量都由于綠原酸處理出現(xiàn)明顯下調(diào);綠原酸處理使超氧化物歧化酶,過敏蛋白Prup1和病程相關蛋白10等具有防御性功能的蛋白表達量出現(xiàn)上調(diào),;碳水化合物代謝相關蛋白質(zhì),包括α-淀粉酶,蔗糖合成酶、UDP-葡萄糖焦磷酸化酶、UDP-糖基轉移酶和α-半乳糖苷酶在油桃果實成熟過程中的表達量也明顯受到綠原酸影響而有所下調(diào)。綠原酸還可以顯著抑制1-氨基環(huán)丙烷-1-羧酸氧化酶(ACO)在油桃果實采后成熟過程的蛋白表達量上升。使用iTRAQ定量蛋白組學技術分析蘋果果肉組織中與蘋果內(nèi)源多酚具有親和作用的蛋白質(zhì)。分析得到157個目標蛋白質(zhì),其中包括與果實成熟衰老相關的脂氧合酶、糖基轉移酶、αα-淀粉酶、淀粉支鏈酶、纖維素合成酶、蔗糖磷酸酶、β-半乳糖苷酶、1-氨基環(huán)丙烷-1-羧酸合成酶(ACS)、ACO和乙烯受體蛋白等。實驗以蘋果果肉切片作為模型來研究綠原酸對蘋果果實衰老的影響。與對照組相比,綠原酸處理顯著降低了蘋果果肉切片乙烯釋放量和呼吸速率,減緩果肉切片硬度和可溶性固形物含量的下降。十二烷基苯磺酸鈉-聚丙烯酰胺凝膠電泳和MALDI-TOF/TOF分析顯示,綠原酸處理顯著降低了蘋果果肉切片中脂肪氧合酶、β-半乳糖苷酶、NADP-ME、UDP-葡萄糖焦磷酸化酶等蛋白的表達量,提高了類甜蛋白的蛋白表達量。在體外實驗中發(fā)現(xiàn),蘋果多酚和綠原酸均可抑制ACS、ACO、α-淀粉酶、脂氧合酶、β-葡萄糖苷酶、β-半乳糖苷糖、聚半乳糖醛酸酶(PG)、果膠甲酯酶(PME)、NADP-ME、過氧化物酶(POD)、多酚氧化酶(PPO)的酶活性,并且綠原酸對這些酶活性的抑制效果與蘋果多酚相比沒有顯著差異。進一步實驗發(fā)現(xiàn),當蘋果果肉切片經(jīng)過綠原酸處理后,其中ACO、ACS、α-淀粉酶、脂氧合酶、β-葡糖糖苷酶、β-半乳糖苷酶、NADP-蘋果酸酶、POD、PPO的酶活性在切片孵育觀察過程中會顯著下降,而PG和PME酶活性沒有受到明顯的影響。
[Abstract]:Phenols are important secondary metabolites in plant body. They have been paid more and more attention to the antioxidation, anti aging, prevention of cardiovascular diseases and the improvement of cognition in human body. However, there are few studies on the effect of phenolic substances on the mature and aging process of postharvest fruits. The contents and results are as follows: compared with the new chlorogenic acid (another main phenolic substance in the fruit flesh of nectarine fruit), chlorogenic acid can be significantly reduced. The release of ethylene in Nectarine pulp sliced, and the content of microchip hardness and soluble solids decreased. Chlorogenic acid could significantly reduce the ethylene release, respiration rate, softening degree, decay rate and malondialdehyde content during storage. At the same time, chlorogenic acid treatment also postponed the red color of nectarine fruit peel during storage. The study also found that chlorogenic acid significantly enhanced the antioxidant activity, free radical scavenging ability and reduction ability of the fruit. The experiment was successfully separated and identified by two dimensional electrophoresis and matrix assisted laser analytical ionization flight two mass spectrometry (MALDI-TOF-TOF/MS). 74 of the fruits of Nectarine were successfully separated and identified. Protein NADP- malate enzyme (NADP-ME), malate dehydrogenase and NAD- dependent malate dehydrogenase, which are involved in fruit respiration, are all down regulated by chlorogenic acid treatment; chlorogenic acid treatment makes superoxide dismutase, allergy protein Prup1 The protein expression of defensive function, such as course related protein 10, was up-regulated, and carbohydrate metabolism related proteins, including alpha amylase, sucrose synthase, UDP- glucose pyrophosphorylase, UDP- glycosyltransferase and alpha galactosidase, were also affected by chlorogenic acid in the fruit ripening of nectarine fruit. The protein expression of 1- amino cyclopropane -1- carboxylic oxidase (ACO) in the Postharvest Ripening of nectarine fruit was also significantly inhibited. The protein content of Apple endogenous polyphenols in apple pulp tissues was analyzed by iTRAQ quantitative proteomics technology. 157 target proteins were obtained, including the analysis of the protein. Lipoxygenase, glycosyltransferase, alpha alpha amylase, amylopectin, cellulose synthetase, sucrose phosphatase, beta galactosidase, 1- aminopropane -1- carboxylic synthetase (ACS), ACO and ethylene receptor protein, etc. were used as a model to study the effect of chlorogenic acid on apple fruit senescence. Compared with the control group, chlorogenic acid treatment significantly reduced the ethylene release and respiration rate of the slices of apple pulp, and slowed down the content of the firmness and soluble solids in the slices of pulp. Twelve alkylbenzene sulfonates polyacrylamide gel electrophoresis and MALDI-TOF/TOF analysis showed that chlorogenic acid treatment significantly reduced the fat in the slices of apple pulp. The expression of lipoxygenase, beta galactosidase, NADP-ME, UDP- glucose pyrophosphorylase and other proteins increased the protein expression of sweet protein. In vitro, apple polyphenols and chlorogenic acids could inhibit ACS, ACO, alpha amylase, lipoxygenase, beta glucosidase, beta galactosidase, poly galacturonase (PG), pectin a, and pectin a The enzyme activity of esterase (PME), NADP-ME, peroxidase (POD) and polyphenol oxidase (PPO), and the inhibitory effect of chlorogenic acid on these enzymes was not significantly different from that of apple polyphenols. Further experiments showed that when the slices of apple pulp were treated with chlorogenic acid, ACO, ACS, alpha amylase, lipoxygenase, beta glucosidase, beta galactozyme, and beta galactosidase were found. The activities of glucosidase, NADP- malic enzyme, POD and PPO decreased significantly during the incubation process, while PG and PME enzyme activities were not significantly affected.
【學位授予單位】：中國農(nóng)業(yè)大學
【學位級別】：博士
【學位授予年份】：2017
【分類號】：TS255.3

【參考文獻】

相關期刊論文 前10條

1 梁敏華;雷建敏;邵佳蓉;蘇新國;楊震峰;陳偉;;UV-C處理對桃果實酚類物質(zhì)代謝和貯藏品質(zhì)的影響[J];核農(nóng)學報;2015年06期

2 楊倩;王丹;常麗麗;孫勇;靳翔;王旭初;;生物質(zhì)譜技術研究進展及其在蛋白質(zhì)組學中的應用[J];中國農(nóng)學通報;2015年01期

3 朱璇;侯媛媛;王英;齊會楠;;水楊酸處理對杏果實呼吸速率和乙烯生物合成的影響[J];食品研究與開發(fā);2013年24期

4 劉穎;郭明曄;白根本;;綠原酸的研究進展[J];中藥材;2012年07期

5 王潔;;多酚-蛋白質(zhì)相互作用的影響因素及其功能特性研究進展[J];河南工業(yè)大學學報(自然科學版);2012年03期

6 徐國前;張振文;郭安鵲;欒麗英;;植物多酚抗逆生態(tài)作用研究進展[J];西北植物學報;2011年02期

7 吳小華;頡敏華;呂建國;李梅;張永茂;李冰峰;;納米SiO_x涂膜對靈武長棗采后品質(zhì)的影響[J];北方園藝;2010年09期

8 杜淑霞;歐仕益;貝惠玲;徐麗;;茶多酚與牛奶蛋白互作對蛋白質(zhì)離體消化率的影響[J];食品與發(fā)酵工業(yè);2010年02期

9 王林纖;戴勇;涂植光;;iTRAQ標記技術與差異蛋白質(zhì)組學的生物標志物研究[J];生命的化學;2010年01期

10 羅磊;郭曉園;;金銀花提取液抗氧化活性研究[J];食品科學;2009年21期

相關博士學位論文 前3條

1 馬琳;殼寡糖復合其他保鮮劑對杏果貯藏品質(zhì)的影響[D];中國農(nóng)業(yè)大學;2015年

2 張鵬;黃瓜果實彎曲性QTL定位及蛋白質(zhì)組差異研究[D];東北農(nóng)業(yè)大學;2009年

3 潘秋紅;果實酸性轉化酶研究[D];中國農(nóng)業(yè)大學;2003年

相關碩士學位論文 前3條

1 羅凱;拮抗酵母結合化學物質(zhì)提高草莓果實采后貯藏性能的研究[D];南京農(nóng)業(yè)大學;2012年

2 王一鳴;桃果實發(fā)育硬核期蛋白質(zhì)雙向電泳方法的建立及應用[D];新疆農(nóng)業(yè)大學;2007年

3 張星海;金銀花中綠原酸提取、分離純化工藝和活性研究[D];浙江大學;2003年

，

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