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目錄

引言……………………………………………………………………………………1

緒論……………………………………………………………………………………2

1.1醋酸菌的性質(zhì)及分類……………………………………………………………2

1.2耐高溫產(chǎn)酸發(fā)酵研究現(xiàn)狀…………………………………………………2

1.3醋酸的應(yīng)用…………………………………………………………………3

1.4菌種篩選及誘變方法………………………………………………………3

1.4.1菌種篩選方法……………………………………………………………3

1.4.2誘變方法…………………………………………………………………4

1.5菌種產(chǎn)物的檢測(cè)方法……………………………………………………………4

1.6本課題研究的主要內(nèi)容及意義…………………………………………………4

第二章 實(shí)驗(yàn)材料與方法……………………………………………………………6

2.1藥品試劑及儀器設(shè)備……………………………………………………………6

2.1.1試劑……………………………………………………………………………6

2.1.2儀器設(shè)備………………………………………………………………………6

2.2菌種來(lái)源…………………………………………………………………………6

2.3培養(yǎng)基……………………………………………………………………………6

2.4菌種篩選…………………………………………………………………………7

2.5菌體生長(zhǎng)曲線的測(cè)定……………………………………………………………7

2.6誘變選育…………………………………………………………………………7

2.6.1誘變選育………………………………………………………………………7

2.6.2計(jì)算菌種致死率………………………………………………………………8

2.7耐熱能力實(shí)驗(yàn)……………………………………………………………………8

2.8正交優(yōu)化…………………………………………………………………………8

第三章  結(jié)果與分析…………………………………………………………………9

3.1顯色劑顯色………………………………………………………………………9

3.2培養(yǎng)基顯色………………………………………………………………………9

3.3單菌落培養(yǎng)………………………………………………………………………11

3.4搖瓶培養(yǎng)情況……………………………………………………………………12

3.5醋酸菌的顯微鏡觀察……………………………………………………………12

3.6菌體濃度和產(chǎn)酸量的測(cè)量………………………………………………………13

3.6.1菌體生長(zhǎng)曲線繪制……………………………………………………………13

3.6.2產(chǎn)酸量曲線……………………………………………………………………14

3.7菌種篩選…………………………………………………………………………15

3.7.1篩選初始菌種…………………………………………………………………15

3.7.2篩選誘變菌株…………………………………………………………………16

3.8正交實(shí)驗(yàn)…………………………………………………………………………17

3.8.1單因素的影響…………………………………………………………………17

3.8.2三因素的正交實(shí)驗(yàn)……………………………………………………………19

3.9篩選耐高溫的菌株………………………………………………………………19

3.9.1初步確定所耐最高溫度………………………………………………………19

3.9.2驗(yàn)證所耐最高溫度……………………………………………………………20

第四章 結(jié)論和展望…………………………………………………………………21

4.1結(jié)論………………………………………………………………………………21

4.2展望………………………………………………………………………………21

致 謝…………………………………………………………………………………23

參考文獻(xiàn)………………………………………………………………………………24

附表A英文參考文獻(xiàn)翻譯

附表B主要參考文獻(xiàn)題錄及摘要

摘    要

醋酸是重要的有機(jī)酸，在醫(yī)藥、食品、日化各個(gè)領(lǐng)路都發(fā)揮重要作用。本課題采用的是生物法來(lái)獲得產(chǎn)醋酸的菌株，探尋獲得最大經(jīng)濟(jì)效益的方法，因此,加強(qiáng)對(duì)醋酸的研究十分必要。本課題具體為研究以酒精為唯一碳源的培養(yǎng)基篩選出產(chǎn)醋酸的菌株。菌種的初篩采用變色圈，產(chǎn)物的檢測(cè)選用酸堿滴定法。

結(jié)合紫外誘變選育提高產(chǎn)酸量，為了篩選出產(chǎn)率高的菌株，下一步需要對(duì)菌種的培養(yǎng)條件進(jìn)行優(yōu)化，篩選出高產(chǎn)的菌株。并對(duì)這些菌株進(jìn)行菌種鑒定。確定最佳培養(yǎng)條件：PH值為6.0，酒精作為唯一碳源的添加量為3%，接種量為5%。篩選出耐高溫的菌株，確定34℃是誘變后菌株所耐最高溫度。

本實(shí)驗(yàn)所篩選菌種經(jīng)強(qiáng)化確認(rèn)后，就可運(yùn)用到工業(yè)生產(chǎn)中；變色圈法對(duì)于篩選醋酸桿菌很有幫助；紫外誘變技術(shù)簡(jiǎn)單，但工作量較大。

關(guān)鍵詞：醋酸桿菌;菌種篩選;紫外誘變;耐高溫

Study on Ultraviolet Mutagenesising and Breeding of High Temperatur Resistant Acetobacter aceti

Abstract

Acetic acid is an important organic acids .It plays avery important roles in medicine，food,daily chemical.In order to obtain the maximum economic efficiency ,what this topic uses is the biological law obtains produces acid. Therefore, it is very essential for strengthen to the acid research. In this research，This topic makes concrete for the research produces the acid take the alcohol as only carbon source culture medium screening the strain. The mold mushroom spawn initially sieves uses the method of color deterioration circle .The product examination selects Acid-base titration method.

Combined with uv mutation breed in improving acid consumption.In order to screen the high production the strain,It needs to carry on the optimization to mold mushroom spawn's raise condition and carry on the mold mushroom spawn appraisal to these strains.Determine the optimum culture conditions as follows:pH value is 6.0,Alcohol as the only carbon source of additives is 3%,quantity is 5%.High temperature resistant strains is screened.34 ℃ is induced by the highest temperature resistant strains after mutagenesis.

the strains of this study should be strengthened before it can be applied to industrial production. the method of color deterioration circle is helpful to screen Acetobacter aceti.The technology of Ultraviolet mutagenesis is simple，but working capacity is very large. 

Key words:Acetobacter aceti;Strains filtration;Ultraviolet mutagenesis;High temperature resistant