陕西师范大学学报（自然科学版）
陝西師範大學學報（自然科學版）
협서사범대학학보（자연과학판）
JOURNAL OF SHAANXI NORMAL UNIVERSITY(NATURAL SCIENCE EDITION)
2010年
5期
72-76
,共5页
王维卓%范代娣%骆艳娥%马晓轩%朱晨辉%郭佳庆
王維卓%範代娣%駱豔娥%馬曉軒%硃晨輝%郭佳慶
왕유탁%범대제%락염아%마효헌%주신휘%곽가경
类人胶原蛋白Ⅱ%重组大肠杆菌%pH两阶段控制%有机酸
類人膠原蛋白Ⅱ%重組大腸桿菌%pH兩階段控製%有機痠
류인효원단백Ⅱ%중조대장간균%pH량계단공제%유궤산
Human-like collagen Ⅱ%recombinant escherichia coli%pH shift control%organic acid
为了实现类人胶原蛋白（HLCⅡ）的规模化生产,对重组大肠杆菌BL21 3.7在30 L发酵罐中发酵生产类人胶原蛋白Ⅱ过程中的关键参数pH进行了系统的研究.将发酵过程中pH控制在6.0~7.8之间,监测pH对细胞生长以及类人胶原蛋白Ⅱ的产量的影响,从而确定出合理的pH控制策略增加类人胶原蛋白Ⅱ的产量.由实验数据可得,在pH7.5条件下细胞浓度达到最大,为68.5 g/L,但HCLⅡ的浓度为8.5 g/L.在pH6.8条件下,HLCⅡ浓度达到最大,为10.5 g/L,但其细胞浓度只有60.9 g/L.同时,对发酵过程中代谢副产物有机酸的含量也进行了监测,随着发酵过程pH值的升高,总有机酸的量也相应的增加,当发酵过程pH控制在7.8时,总有机酸达到最大,为15.61 g/L.为了提高目标产物的产量,采用pH两阶段控制策略控制发酵,最终类人胶原蛋白Ⅱ浓度可达到11.3 g/L,较恒pH条件下提高了7.61%.
為瞭實現類人膠原蛋白（HLCⅡ）的規模化生產,對重組大腸桿菌BL21 3.7在30 L髮酵罐中髮酵生產類人膠原蛋白Ⅱ過程中的關鍵參數pH進行瞭繫統的研究.將髮酵過程中pH控製在6.0~7.8之間,鑑測pH對細胞生長以及類人膠原蛋白Ⅱ的產量的影響,從而確定齣閤理的pH控製策略增加類人膠原蛋白Ⅱ的產量.由實驗數據可得,在pH7.5條件下細胞濃度達到最大,為68.5 g/L,但HCLⅡ的濃度為8.5 g/L.在pH6.8條件下,HLCⅡ濃度達到最大,為10.5 g/L,但其細胞濃度隻有60.9 g/L.同時,對髮酵過程中代謝副產物有機痠的含量也進行瞭鑑測,隨著髮酵過程pH值的升高,總有機痠的量也相應的增加,噹髮酵過程pH控製在7.8時,總有機痠達到最大,為15.61 g/L.為瞭提高目標產物的產量,採用pH兩階段控製策略控製髮酵,最終類人膠原蛋白Ⅱ濃度可達到11.3 g/L,較恆pH條件下提高瞭7.61%.
위료실현류인효원단백（HLCⅡ）적규모화생산,대중조대장간균BL21 3.7재30 L발효관중발효생산류인효원단백Ⅱ과정중적관건삼수pH진행료계통적연구.장발효과정중pH공제재6.0~7.8지간,감측pH대세포생장이급류인효원단백Ⅱ적산량적영향,종이학정출합리적pH공제책략증가류인효원단백Ⅱ적산량.유실험수거가득,재pH7.5조건하세포농도체도최대,위68.5 g/L,단HCLⅡ적농도위8.5 g/L.재pH6.8조건하,HLCⅡ농도체도최대,위10.5 g/L,단기세포농도지유60.9 g/L.동시,대발효과정중대사부산물유궤산적함량야진행료감측,수착발효과정pH치적승고,총유궤산적량야상응적증가,당발효과정pH공제재7.8시,총유궤산체도최대,위15.61 g/L.위료제고목표산물적산량,채용pH량계단공제책략공제발효,최종류인효원단백Ⅱ농도가체도11.3 g/L,교항pH조건하제고료7.61%.
To achieve large-scale production of HLC Ⅱ,the key parameter pH in production process of human-like collagen by recombinant Escherichia coli BL21 3.7 was systematicly investigated in a 30 L fermentation tank with triplex turbine impeller.The effect of pH ranging from 6.0 to 7.8 on cell content and HLC Ⅱ production during fermentation process was studied,and an optimal pH strategy was determined for increase of HLC Ⅱ yield.From experimental data,the cell growth and collagen synthesis were significantly affected by pH.An optimal pH of 7.5 for cell growth was obtained with cell density at 68.5 g/L,the concentration of HLC Ⅱ was 8.5 g/L;while an optimal pH of 6.8 for HLC Ⅱ accumulation was obtained with concentration of HLC Ⅱ at 10.5 g/L,and cell density was 60.9 g/L.Additionally,the total organic acid of fermentation process was monitored.The concentration of organic acid increased with pH,and the highest concentration of organic acid,15.61 g/L,was observed when pH was kept constant at 7.8 during the process of fermentation.Based on the results,a pH shift method was developed to gain higher concentration of HCL Ⅱ which reached to 11.3 g/L,increased by 7.61% as compared with constant pH fermentation.
