CIESC Journal ›› 2022, Vol. 73 ›› Issue (10): 4311-4323.DOI: 10.11949/0438-1157.20220838
• Reviews and monographs • Previous Articles Next Articles
Xinhui WANG1,2,3(), Ying WANG1,2, Mingdong YAO1,2,3(), Wenhai XIAO1,2
Received:
2022-06-15
Revised:
2022-07-27
Online:
2022-11-02
Published:
2022-10-05
Contact:
Mingdong YAO
王欣慧1,2,3(), 王颖1,2, 姚明东1,2,3(), 肖文海1,2
通讯作者:
姚明东
作者简介:
王欣慧(1998—),女,硕士研究生,1575799416@qq.com
基金资助:
CLC Number:
Xinhui WANG, Ying WANG, Mingdong YAO, Wenhai XIAO. Research progress of vitamin A biosynthesis[J]. CIESC Journal, 2022, 73(10): 4311-4323.
王欣慧, 王颖, 姚明东, 肖文海. 维生素A生物合成的研究进展[J]. 化工学报, 2022, 73(10): 4311-4323.
公司 | 产地 | 上游中间体 | 生产工艺 | 产能/(t/a) |
---|---|---|---|---|
新和成 | 中国 | 自配 | Roche[ | 10000 |
帝斯曼 | 荷兰等 | 外购 | Roche | 7500 |
巴斯夫 | 德国等 | 自配 | BASF[ | 6000 |
浙江医药 | 中国 | 外购 | BASF | 5600 |
安迪苏 | 法国等 | 外购 | BASF | 5000 |
金达威 | 中国 | 外购 | Roche | 2900 |
Table 1 Global major manufacturers of vitamin A
公司 | 产地 | 上游中间体 | 生产工艺 | 产能/(t/a) |
---|---|---|---|---|
新和成 | 中国 | 自配 | Roche[ | 10000 |
帝斯曼 | 荷兰等 | 外购 | Roche | 7500 |
巴斯夫 | 德国等 | 自配 | BASF[ | 6000 |
浙江医药 | 中国 | 外购 | BASF | 5600 |
安迪苏 | 法国等 | 外购 | BASF | 5000 |
金达威 | 中国 | 外购 | Roche | 2900 |
底盘细胞 | 工程化手段 | 产量 | 文献 |
---|---|---|---|
大肠杆菌 | 表达家鼠来源的β-胡萝卜素15,15′-氧化酶; 优化最佳反应条件:pH、温度、底物浓度和表面活性剂 | 视黄醛产量:72 mg/L | [ |
大肠杆菌 | 选择密码子优化后的β-胡萝卜素15,15′-氧化酶; 上调dxs,优化内源MEP途径; 引入外源性MVA途径,提供IPP和DMAPP; 筛选大肠菌株:MG1655、DH5a、X11-BLUE、S17-1和BL21; 发酵条件优化:氧浓度、碳源和温度 | 视黄醛产量:136 mg/L | [ |
大肠杆菌 | 引入两个异源基因blhSR和RALDH2; 删除内源编码醛还原酶的ybbO基因; 采用mRNA稳定区工程方法; 优化维生素A产生菌的培养条件:温度、pH和氧浓度 | 5 L生物反应器分批发酵;维甲酸产量: (8.20±0.05) mg/L | [ |
大肠杆菌 | 筛选了编码β-胡萝卜素裂解酶的基因:blh、brp和bcox; 采用mRNA稳定区工程提高blh催化效率; 共同表达合成视黄基棕榈酸酯路径基因LRAT和CRBP | 分批补料发酵培养;视黄基棕榈酸酯产量为(69.96±2.64) mg/L | [ |
大肠杆菌 | 优化发酵条件:温度、pH、搅拌速度; 筛选最佳表面活性剂 | 视黄醛产量:600 mg/L | [ |
大肠杆菌 | 过表达内源基因ybbO,增强醛→醇转化; 敲除编码O-乙酰转移酶的基因cat | 视黄醛、视黄醇、视黄醇醋酸酯产量的比例为6%、88%、6% | [ |
酿酒酵母 | 采用以木糖为碳源的酵母菌株; 采用十二烷和橄榄油两相原位萃取 | 3 L生物反应器分批补料;视黄醇、视黄醛产量为1256、2094 mg/L | [ |
酿酒酵母 | 引入人来源RDH12和乳球菌noxE; 优化发酵条件,以木糖为碳源,十二烷为萃取剂; 筛选最适培养和储存条件,包括温度、光照和抗氧化剂 | 250 ml摇瓶发酵;视黄醇产量:123.1 mg/L | [ |
酿酒酵母 | 过表达酵母内源ENV9和截短的Hmg1; 发酵添加1.44 mmol/L Fe2+以维持blh酶活性 | 视黄醇产量:443.43 mg/L | [ |
Table 2 Progress in metabolic engineering transformation related to vitamin A
底盘细胞 | 工程化手段 | 产量 | 文献 |
---|---|---|---|
大肠杆菌 | 表达家鼠来源的β-胡萝卜素15,15′-氧化酶; 优化最佳反应条件:pH、温度、底物浓度和表面活性剂 | 视黄醛产量:72 mg/L | [ |
大肠杆菌 | 选择密码子优化后的β-胡萝卜素15,15′-氧化酶; 上调dxs,优化内源MEP途径; 引入外源性MVA途径,提供IPP和DMAPP; 筛选大肠菌株:MG1655、DH5a、X11-BLUE、S17-1和BL21; 发酵条件优化:氧浓度、碳源和温度 | 视黄醛产量:136 mg/L | [ |
大肠杆菌 | 引入两个异源基因blhSR和RALDH2; 删除内源编码醛还原酶的ybbO基因; 采用mRNA稳定区工程方法; 优化维生素A产生菌的培养条件:温度、pH和氧浓度 | 5 L生物反应器分批发酵;维甲酸产量: (8.20±0.05) mg/L | [ |
大肠杆菌 | 筛选了编码β-胡萝卜素裂解酶的基因:blh、brp和bcox; 采用mRNA稳定区工程提高blh催化效率; 共同表达合成视黄基棕榈酸酯路径基因LRAT和CRBP | 分批补料发酵培养;视黄基棕榈酸酯产量为(69.96±2.64) mg/L | [ |
大肠杆菌 | 优化发酵条件:温度、pH、搅拌速度; 筛选最佳表面活性剂 | 视黄醛产量:600 mg/L | [ |
大肠杆菌 | 过表达内源基因ybbO,增强醛→醇转化; 敲除编码O-乙酰转移酶的基因cat | 视黄醛、视黄醇、视黄醇醋酸酯产量的比例为6%、88%、6% | [ |
酿酒酵母 | 采用以木糖为碳源的酵母菌株; 采用十二烷和橄榄油两相原位萃取 | 3 L生物反应器分批补料;视黄醇、视黄醛产量为1256、2094 mg/L | [ |
酿酒酵母 | 引入人来源RDH12和乳球菌noxE; 优化发酵条件,以木糖为碳源,十二烷为萃取剂; 筛选最适培养和储存条件,包括温度、光照和抗氧化剂 | 250 ml摇瓶发酵;视黄醇产量:123.1 mg/L | [ |
酿酒酵母 | 过表达酵母内源ENV9和截短的Hmg1; 发酵添加1.44 mmol/L Fe2+以维持blh酶活性 | 视黄醇产量:443.43 mg/L | [ |
改造策略 | 具体改造信息 | 结果 | 文献 |
---|---|---|---|
碳源 | 葡萄糖、木糖 | 20.74 mg/L维生素A(44 g/L木糖) | [ |
温度、pH | 温度:24~33℃ pH:5.5~8.0 | 视黄醛产量最大(pH 7.0,30℃), 细胞量达到最大(27℃) | [ |
表面活性剂 | Span20、Span40、Span60、Span80、 Tween20、Tween40、Tween80、Brij58、Triton X-100 | 细胞量最大, 细胞积累量为15.2 g/L(10 g/L Span80) | [ |
搅拌速率 | 400、500、600、700、800 r/min | 视黄醛产量最大为600 mg/L(600 r/min) | [ |
萃取剂 | 十二烷、大豆油和橄榄油 | 十二烷效果最佳 | [ |
光 | 光照条件、黑暗条件 | 视黄醇浓度最高为124.6 mg/L(黑暗) | [ |
Table 3 The downstream transformation strategy of vitamin A production
改造策略 | 具体改造信息 | 结果 | 文献 |
---|---|---|---|
碳源 | 葡萄糖、木糖 | 20.74 mg/L维生素A(44 g/L木糖) | [ |
温度、pH | 温度:24~33℃ pH:5.5~8.0 | 视黄醛产量最大(pH 7.0,30℃), 细胞量达到最大(27℃) | [ |
表面活性剂 | Span20、Span40、Span60、Span80、 Tween20、Tween40、Tween80、Brij58、Triton X-100 | 细胞量最大, 细胞积累量为15.2 g/L(10 g/L Span80) | [ |
搅拌速率 | 400、500、600、700、800 r/min | 视黄醛产量最大为600 mg/L(600 r/min) | [ |
萃取剂 | 十二烷、大豆油和橄榄油 | 十二烷效果最佳 | [ |
光 | 光照条件、黑暗条件 | 视黄醇浓度最高为124.6 mg/L(黑暗) | [ |
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