產(chǎn)β-苯丙氨酸變位酶基因工程菌的構(gòu)建及其發(fā)酵條件優(yōu)化
發(fā)布時(shí)間：2019-05-14 12:26
【摘要】：構(gòu)建β-苯丙氨酸變位酶基因表達(dá)重組質(zhì)粒pET-28apam,并轉(zhuǎn)化大腸桿菌BL21,獲得β-苯丙氨酸變位酶基因重組菌�？疾炫囵B(yǎng)基初始pH、搖床轉(zhuǎn)速、種齡、發(fā)酵時(shí)間以及裝液量5個(gè)因素對β-苯丙氨酸變位酶基因工程菌產(chǎn)酶活性的影響,確定較佳發(fā)酵條件為:培養(yǎng)基初始pH 7.0、搖床轉(zhuǎn)速200r/min、發(fā)酵時(shí)間24h、裝液量50mL/500mL、種齡10h。采用Box-Behnken試驗(yàn)設(shè)計(jì),選擇了種齡、初始pH、發(fā)酵時(shí)間3個(gè)對β-苯丙氨酸變位酶活力影響較大的因素進(jìn)行響應(yīng)面優(yōu)化。優(yōu)化結(jié)果為:在種齡12.8h、初始pH 6.7、發(fā)酵時(shí)間25.2h條件下,PAM活力達(dá)到11.15 U/mL,比優(yōu)化前酶活4.87U/mL提高了128.9%。
[Abstract]:The recombinant plasmid pET-28apam, was constructed and transformed into E. coli BL21, to obtain the recombinant strain of 尾-phenylalanine translocation gene. The effects of initial pH, shaking speed, seed age, fermentation time and liquid loading on the enzyme production activity of 尾-phenylalanine translocation genetic engineering bacteria were investigated. The optimum fermentation conditions were as follows: the initial pH 7.0, The rotating speed of shaking table was 200 r / min, the fermentation time was 24 h, the amount of liquid was 50 mL / 500 mL, and the seed age was 10 h. Using Box-Behnken design, three factors, seed age and initial pH, fermentation time, which had great influence on the activity of 尾-phenylalanine translocation enzyme, were selected to optimize the response surface. The results showed that under the conditions of seed age 12.8 h, initial pH 6.7 and fermentation time 25.2 h, the activity of PAM reached 11.15 U / mL, which was 128.9% higher than that before optimization.
【作者單位】： 安徽工程大學(xué)生物與化學(xué)工程學(xué)院;
【基金】：國家自然科學(xué)基金面上項(xiàng)目(編號:31671797) 安徽省高等學(xué)校省級自然科學(xué)研究重點(diǎn)項(xiàng)目(編號:KJ2016A801) 國家級大學(xué)生創(chuàng)新創(chuàng)業(yè)訓(xùn)練計(jì)劃項(xiàng)目(編號:201510363072)
【分類號】：TS201.3
本文編號：2476714
[Abstract]:The recombinant plasmid pET-28apam, was constructed and transformed into E. coli BL21, to obtain the recombinant strain of 尾-phenylalanine translocation gene. The effects of initial pH, shaking speed, seed age, fermentation time and liquid loading on the enzyme production activity of 尾-phenylalanine translocation genetic engineering bacteria were investigated. The optimum fermentation conditions were as follows: the initial pH 7.0, The rotating speed of shaking table was 200 r / min, the fermentation time was 24 h, the amount of liquid was 50 mL / 500 mL, and the seed age was 10 h. Using Box-Behnken design, three factors, seed age and initial pH, fermentation time, which had great influence on the activity of 尾-phenylalanine translocation enzyme, were selected to optimize the response surface. The results showed that under the conditions of seed age 12.8 h, initial pH 6.7 and fermentation time 25.2 h, the activity of PAM reached 11.15 U / mL, which was 128.9% higher than that before optimization.
【作者單位】： 安徽工程大學(xué)生物與化學(xué)工程學(xué)院;
【基金】：國家自然科學(xué)基金面上項(xiàng)目(編號:31671797) 安徽省高等學(xué)校省級自然科學(xué)研究重點(diǎn)項(xiàng)目(編號:KJ2016A801) 國家級大學(xué)生創(chuàng)新創(chuàng)業(yè)訓(xùn)練計(jì)劃項(xiàng)目(編號:201510363072)
【分類號】：TS201.3
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