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木質素酶產生真菌的分離純化及鑒定

時間:2019-04-07 20:35來源:畢業論文
篩選木質素酶產生菌株。從學校腐木以及柳樹、松樹、葡萄樹等樹皮中分離純化出能夠降解秸稈中木質素的真菌,并測定漆酶和木質素過氧化物酶的活性及菌種進行鑒定,從而篩選出高

摘要:作為一種潛在的可再生資源,纖維素和木質素具有很大的利用價值。但是因為纖維素經常會與半纖維素以及木質素等一些較難分解物質結合在一起,因此在利用秸稈等含纖維素豐富的有機質之前,就要將其與木質素和半纖維素分開,從而使纖維素暴露出來,以便更好地與降解酶接觸;相對于木質素而言,半纖維素降解較為容易,因此秸稈中纖維素利用的核心就是木質素降解問題,通過微生物來完成木質素的去處是具有潛在價值的熱點之一。本實驗目的就是篩選木質素酶產生菌株。從學校腐木以及柳樹、松樹、葡萄樹等樹皮中分離純化出能夠降解秸稈中木質素的真菌,并測定漆酶和木質素過氧化物酶的活性及菌種進行鑒定,從而篩選出高產漆酶和木質素過氧化物酶的真菌。為篩選能高效降解木質素的菌株,從11份采集的朽木與樹皮樣品中分離純化獲得36株菌,將純化獲得的菌株分別接入到PDA-愈創木酚和PDA-苯胺藍培養基上進行初篩,觀察其顏色變化,根據變色圈的大小和深淺篩選漆酶產生菌,根據褪色圈的大小篩選木質素過氧化物酶產生菌,共篩選出11株產漆酶或木質素過氧化物酶的菌株,將這11株菌接入液體產酶培養基中進行液體靜止培養并測定木質素酶活力,最終獲得1株高產木質素酶的真菌菌株L7。其所產木質素過氧化物酶活力達到207.1U/L;在液體培養基中,漆酶測定值較小,受誤差影響較大,特采用固體培養基測漆酶,將板子上顯色明顯的菌株接種到固體培養基上,測定漆酶活性,最終篩選一株高產漆酶菌株1-1,其漆酶活力達到1033.1U/g。34365
畢業論文關鍵字:木質素; 漆酶; 木質素過氧化物酶; 愈創木酚; 苯胺藍
Separation Purification and Identification of Lignin Enzyme Producing Fungi

源¥自%六^^維*論-文+網=www.aftnzs.live


Abstract: As a potentially renewable resource, cellulose and lignin have great value. But the cellulose is easy to combine with hemicellulose, lignin and other matters which are hard to break down, so it must be released from lignocellulose and hemicellulose in the parcel before cellulose was used. And hemicellulose can be degraded easily, therefore, the key of cellulose degradation is the degradation of lignin, so the use of microbial degradation of lignin and cellulose has been one of the attention. The objective of this experiment is screening strains which can produce lignin enzyme.  In order to separate and purify lignin-degradation fungi, fungi were screened from the rotten wood and the barks of willow, pine and grape trees. The activity of laccase and lignin peroxidase were determined to screen strains which can produce high yield laccase and lignin peroxidase. 36 strains was purified from the rotten wood and barks, with the purpose of screening high efficient lignin-degradation fungi, these purified strains were inoculated in the PDA-guaiacol plate and PDA-aniline blue plate to observe the color change, screening laccase producing fungi according to the size and depth of color ring and screening lignin peroxidase producing fungi according to the size of fade circle. 11 strains were screened and then inoculated in the liquid culture medium with static culture to determine the lignin enzyme activity, results showed that L7 was the most excellent strain with the higher lignin-degrading enzymes activities for its lignin peroxidase were up to 207.1U/L. In the liquid culture medium, the activity of laccase is smaller, and the numerical value is influenced bigger by error, so using the solid medium measure laccase activity, results showed that 1-1 was the most excellent strain with the higher laccase enzymes activities for its laccase activity were up to 1033.1U/g.
   Key words: lignin; laccase; lignin peroxidase; guaiacol; aniline blue
目錄
引言    1

源¥自%六^^維*論-文+網=www.aftnzs.live


1實驗材料與方法    3
1.1樣本來源    3 木質素酶產生真菌的分離純化及鑒定:http://www.aftnzs.live/shengwu/20190407/31838.html
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