【正文】
e t i m e ( h )HA concentration (g/L)0 5 10 15 200204060C u l t u r e t i m e ( h )Lactic acid concentration (g/L)( a ) ( b )( c ) ( d )HA氧化還原降解對(duì) HA發(fā)酵的影響 添加 H2O2/抗壞血酸對(duì)流體力學(xué)影響 添加 H2O2,Vc 31 Transglutaminase (TGase, Protein glutamine γglutamyltransferase, EC ) 舉例 4: Transglutaminase: Activation Mechanism and Fermentation Optimization ? 通過 Nε(γ谷酰胺 ) 賴氨酸鍵交聯(lián)蛋白質(zhì) ? 催化蛋白質(zhì)中 Gln的 γ羧酰胺基與伯胺間發(fā)生轉(zhuǎn)移反應(yīng) ? 蛋白質(zhì)脫酰胺作用 ? 通過形成脂鍵共價(jià)連接蛋白質(zhì)和脂肪酸的長鏈 ω羥基 ? 目前唯一商業(yè)化并大規(guī)模生產(chǎn)的可在蛋白質(zhì)間形成共價(jià)交聯(lián)的酶制劑 32 Activation Mechanism of Transglutaminase PreProTGase TA PCT AB 抑制因子復(fù)合物沉淀+活化 TGaseCT AB+TA P 抑制因子復(fù)合物從細(xì)胞分泌抑制因子TA P+細(xì)胞復(fù)合物沉淀抑制因子復(fù)抑制因子復(fù)抑制因子復(fù)從細(xì)胞分泌從細(xì)胞分泌抑制因子 Secreted from cells folding ProTGase Inhibitor (TAPI) Inhibition Proteaseinhibitor plexs activation Serine Protease MetalloProtease TGase Proregion (AAs) + 舉例 4: Transglutaminase: Activation Mechanism and Fermentation Optimization 33 根據(jù)該 TGase活化模型:在發(fā)酵過程的產(chǎn)酶初期 : *添加 1000 U/mL的胰蛋白酶,使發(fā)酵過程酶活提高 % *添加 10 mg/mL的 CTAB,使發(fā)酵過程酶活提高 % 舉例 4: Transglutaminase: Activation Mechanism and Fermentation Optimization 34 Gl u cos e G6 P F 6 P GAP G3 P P EP P y r O AA R 5 P Hi s S er C y s T rp P h e T y r Al a V al L e u As p Gl y L y s Th r M et As n I l e AcC o A C it AKG Gl u P ro Ar g Gl n A m in o acid s for o th er p r o tei n s A m in o acid s for TG ase Hi s C y s S er Gl y T rp P h e T y r Al a V al L e u As p As n M et Th r L y s I l e Gl n Ar g P ro Gl u Determine of culture medium by analysis of amino acid metabolism using mass balances Amino acid metabolic anylysis of Streptomyces Mobaraense Twostage pH and agitation speed control strategy for TGase fermentation. DO1 DO2 DO2 DO1 pH1 pH2 舉例 4: Transglutaminase: Activation Mechanism and Fermentation Optimization pH1:。 pH2: DO1:轉(zhuǎn)速 300 rpm。 DO2:轉(zhuǎn)速 200 rpm 35 小 結(jié) ? 對(duì)微生物發(fā)酵過程進(jìn)行解析(生理、動(dòng)力學(xué)、代謝網(wǎng)絡(luò)) ? 根據(jù)不同發(fā)酵過程的特點(diǎn),提出相應(yīng)的優(yōu)化控制策略 ?實(shí)現(xiàn)微生物發(fā)酵過程高產(chǎn)量、高產(chǎn)率和高生產(chǎn)強(qiáng)度的有機(jī)統(tǒng)一 36 謝謝!