論文谷氨酰胺轉(zhuǎn)胺酶對小麥蛋白質(zhì)和淀粉催化研究

1、TG論文：谷氨酰胺轉(zhuǎn)胺酶對小麥蛋白質(zhì)和淀粉催化研究【中文摘要】谷氨酰胺轉(zhuǎn)胺酶(TGase,簡稱TG)是一種催化?；D(zhuǎn)移反應的轉(zhuǎn)移酶,作為一種新型、安全的酶制劑,在國外已經(jīng)歷了十幾年的研究,進行了大量的試驗并逐漸應用于面粉制品生產(chǎn)中,而國內(nèi)這方面的研究較少。目前,國內(nèi)外對谷氨酰轉(zhuǎn)胺酶在面制品中的應用研究主要集中在制粉和面制品加工兩方面:在小麥制粉的潤麥、混合等工序中使用TG可改善面粉品質(zhì),延長保質(zhì)期,降低面粉致敏性;在焙烤食品如面包、曲奇餅、蛋糕等加工中使用,可改善食品品質(zhì)。因此,利用TG的共價交聯(lián)化學特性對面粉品質(zhì)進行改善,在面粉制品的加工中具有重大的意義。本文針對TG對蛋白質(zhì)的交聯(lián)催化反應,

2、主要研究了TG對小麥蛋白質(zhì)和淀粉顆粒組分內(nèi)及組分間的催化機理,以及TG對小麥面團結(jié)構特性的影響。為面粉加工品的發(fā)展奠定了理論基礎。主要獲得了以下結(jié)論:(1)采用組分分離方法,獲得了小麥清蛋白、球蛋白、醇溶蛋白和麥谷蛋白等各個蛋白組分和淀粉顆粒,通過電子顯微鏡觀察、可溶性蛋白含量的檢測以及SDS-PAGE電泳分析,研究了TG對這些蛋白質(zhì)的單一組分、兩種蛋白組分、三種蛋白組分和四種蛋白組分內(nèi)部及相互間作用的影響,重點考察了作用過程中可溶性蛋白含量和蛋白質(zhì)電泳條帶的變化;用掃描電子顯微鏡觀察了酶作用條件下,每種單蛋白組分、兩種蛋白組分、三種蛋白組分及四種蛋白組分與淀粉顆粒表面之間作用的影響。結(jié)果發(fā)現(xiàn)

3、,TG可以催化小麥中清蛋白、球蛋白和醇溶蛋白組分的分子內(nèi)和分子間的作用;對球蛋白的催化作用大于清蛋白,但兩者均小于醇溶蛋白;清蛋白對TG催化其它蛋白組分的分子內(nèi)結(jié)合有促進作用;球蛋白對TG催化醇溶蛋白分子內(nèi)結(jié)合都有一定的抑制作用。(2)試驗以小麥面團加酶前后,從面團的微觀結(jié)構、蛋白質(zhì)組分變化等方面研究了TG對小麥中蛋白組分間結(jié)合作用的影響。結(jié)果發(fā)現(xiàn):TG可以增強小麥面團中面筋結(jié)構的強度;催化小麥清蛋白和球蛋白的相互作用;對小麥球蛋白的催化作用大于小麥清蛋白?！居⑽恼縏ransglutaminase(TG) as a new, safe enzyme, which has been rese

4、arched about ten years in foreign country. A series of tests had been carried out and it has gradually applied to the production of flour products, but there is less research about this area in domestic. The present researches of Transglutaminase focused on the two aspects of milling and pasta proce

5、ssing : the use of TG can improve flour quality, extend shelf life and reduce the allergenicity of wheat flour in the processes of tempering and mixing wheat flour, it can also rebound wheat quality when TG was used to hand Worm-eaten wheat and improve food quality of baked goods such as bread, cook

6、ies, cakes and so on. Therefore, the use of covalent crosslinking properties of Transglutaminase can improve the flour quality, which has a great significance and foreground in the processing of flour products.In this paper, the catalytic mechanism of TG in and between the components of wheat protei

7、n and starch granules was researched, meanwhile, the effort of group structure of wheat and buckwheat noodles was discussed, all above have laid a theoretical foundation for the development of flour processing.Through this research, the main conclusions were as following:(1)Based on the method of co

8、mponents separation,wheat albumin, globulin, gliadin, glutenin and starch granules were obtained, the effect of transglutaminase on the intra- and interaction of these isolated compounds was studied in the paper. The reaction was studied at the levels of single, two, three and four kinds of protein

9、compounds and starch granules. Before and after the enzymatic reaction, content and molecular weight were analyzed according to the soluble protein content and SDS-PAGE analysis, and microstructure of the starch granules was also observed by using EMS. The results showed that TG could catalyze the i

10、nteractions inner a same protein or between the different proteins including albumin, globulin, and gliadin. The effect of TG was greater on globulin than on albumin, and the greatest on gliadin among the three proteins. Albumin showed a negative effect on TG catalyzing the interaction of globulin a

11、nd gliadin, while globulin showed a negative effect on gliadin.(2)Meanwhile, The Soba and the mixture of buckwheat flour and wheat were used as the control group to research the effect of transglutaminase (TGase) on the interaction among the buckwheat and wheat protein compounds in the aspects of mi

12、crostructure, soluble protein content, and protein compounds. Results showed that TG strengthened the structure of gluten net in dough of wheat, buckwheat and mixture of the both. TG also could catalyze the interaction of albumin and globulin of wheat and buckwheat. As to buckwheat, the effect of TG

13、 was greater on albumin than on globulin, however, for wheat, it was reverse. TG showed greater effect on the mixture of buckwheat albumin and globulin than wheat albumin and globulin.【關鍵詞】TG 清蛋白 球蛋白 醇溶蛋白 麥谷蛋白【英文關鍵詞】Transglutaminase Albumin Globulin Giadin Glutenin【目錄】 小麥淀粉組成及特性 13-14 1.2.2 小麥淀粉對主要面

14、制品的品質(zhì)影響 14-15 1.3 小麥面團特性 15-16 1.3.1 面團的結(jié)構 15 1.3.2 面團的化學鍵 15-16 1.4 TG 研究概況 16-19 1.4.1 TG 種類及特性 16 1.4.2 TG 的催化機理研究 16-18 1.4.3 TG 對谷物面團、蛋白和淀粉的催化作用研究 18-19 1.5 TG 在面制品的應用中存在的問題 19 1.6 研究的目的和意義 19-20 1.7 本文主要的研究內(nèi)容 20-21 1.7.1 研究內(nèi)容 20 1.7.2 技術路線 20-21 第二章 材料、試劑與試驗方法 21-26 2.1 材料、試劑與儀器 21 2.1.1 材料與試劑

15、 21 2.1.2 主要儀器與設備 21 2.2 試驗方法 21-26 2.2.1 蛋白質(zhì)組分的分離 21 2.2.2 淀粉組分的分離 21-22 2.2.3 可溶性蛋白濃度的測定 22 2.2.4 SDS-PAGE 電泳 22-23 2.2.5 顯微結(jié)構觀察 23 2.2.6 TG 對小麥蛋白質(zhì)和淀粉組分內(nèi)催化作用研究 23-24 2.2.7 谷氨酰胺轉(zhuǎn)氨酶對小麥面團內(nèi)部結(jié)構的影響 24-26 第三章 試驗結(jié)果與分析 26-39 3.1 酶促反應速度檢測時間的確定 26-27 3.2 TG 對蛋白質(zhì)組分的催化作用 27-31 3.2.1 TG 對各蛋白組分的酶促反應速度 27 3.2.2 谷氨酰胺轉(zhuǎn)氨酶對各蛋白組分相互作用的影響 27-30 3.2.3 TG 對小麥非麥谷蛋白組分內(nèi)和組分間的催化作用 30-31 3.3. TG 對小麥蛋白質(zhì)和淀粉組分間催化