Synthetic Biology Journal ›› 2022, Vol. 3 ›› Issue (5): 1031-1059.DOI: 10.12211/2096-8280.2022-014
• Invited Review • Previous Articles
Zixuan YOU1,2,3, Feng LI1,2,3, Hao SONG1,2,3
Received:
2022-03-01
Revised:
2022-04-19
Online:
2022-11-16
Published:
2022-10-31
Contact:
Feng LI, Hao SONG
由紫暄1,2,3, 李锋1,2,3, 宋浩1,2,3
通讯作者:
李锋,宋浩
作者简介:
由紫暄(1997—),女,硕士研究生,研究方向为电能细胞的合成生物学研究。基金资助:
CLC Number:
Zixuan YOU, Feng LI, Hao SONG. Design and construction of electroactive cells by synthetic biology strategies[J]. Synthetic Biology Journal, 2022, 3(5): 1031-1059.
由紫暄, 李锋, 宋浩. 电能细胞的合成生物学设计构建[J]. 合成生物学, 2022, 3(5): 1031-1059.
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URL: https://synbioj.cip.com.cn/EN/10.12211/2096-8280.2022-014
Fig. 1 Bi-directional electron transfer mechanism(Left) Direct electron transfer via cytochromes and conductive nanowires; (Right) Electron shuttles mediated electron transfer
电能细胞 | 合成生物学策略 | 工程结果 |
---|---|---|
底物代谢效率的合成生物学改造 | ||
S.oneidensis | 强化乳酸利用关键因子CRP表达,增强菌株对乳酸的利用能力[ | 最大电流密度约40 mA/m2 |
异源表达视紫红质基因,通过光驱动改变膜电位,增强乳酸的摄取[ | 平均电流密度达0.39 A/m2 | |
异源表达葡萄糖转运基因和葡萄糖激酶基因,增强菌株葡萄糖代谢能力[ | 最大电流输出0.085 mA | |
异源表达木糖转运蛋白、异构酶等构建木糖代谢途径[ | 最大功率密度2.1 mW/m2 | |
异源表达乙酸辅酶A基因(ato1,ato2),强化柠檬酸合酶基因gtlA,构建乙酸代谢路径[ | 最大功率密度8.3 mW/m2 | |
胞内可释放电子池的合成生物学改造 | ||
P.aeruginosa | 过表达NAD+合成酶基因nadE,扩充胞内NAD(H/+)总量[ | 最大功率密度4.0 mW/m2 |
S.oneidensis | 过表达ycel、pncB、nadM、nadD*、nadE*基因,增强NAD+的生物合成[ | 最大功率密度162.8 mW/m2 |
过表达gapA2、pflB、fdh*、mdh基因,强化NADH的合成[ | 最大功率密度105.8 mW/m2 | |
E.coli | 破坏乳酸脱氢酶基因ldhA,释放储存在胞内中间代谢物的电子[ | 最大功率密度3.0 mW/m2 |
过表达非质子泵NADH脱氢酶NDH-2,使更多电子通过氧化呼吸链流入EET途径[ | 输出电流4.7 μA | |
构建C3N代谢通路,获得高效的NAD+代谢途径[ | NAD(H)浓度约6 mmol/L | |
胞外电子传递的合成生物学改造 | ||
E.coli | 通过引入S.oneidensis的MtrCAB色素系统使得E.coli具备胞外电子传递能力[ | Fe(Ⅲ)还原速率达83 μmol/(L·d) |
构建S.oneidensis和E.coli细胞色素融合表达系统[ | 电流密度达到25.32 mA/m2 | |
异源表达PCA合成路径phzA1B1C1D1E1F1G1基因簇,以提高间接胞外电子传递速率[ | 最大功率密度为181.1 mW/m2 | |
S.oneidensis | 过表达内膜细胞色素CymA,加速电子传递至末端周质还原酶[ | 功率达到0.13 mW |
敲除周质色素蛋白napB、fccA、tsdB并过表达细胞色素cctA,降低周质细胞色素复杂性[ | 最高功率密度达到436.5 mW/m2 | |
异源表达来自B.subtilis的核黄素合成基因簇ribADEHC,以提高菌株的胞外电子传递速率[ | 最大功率输出为233.0 mW/m2 | |
强化核黄素表达,结合孔蛋白增强分泌,添加材料rGO,构建三维杂合生物膜系统[ | 最大功率输出达2630 mW/m2 | |
异源构建PCA合成路径和ombB-omaB-omcB-omcS直接电子传递链[ | 最大电流密度达310.2 mA/m2 | |
P.aeruginosa | 通过截断PaPilA C端并增加N端 α-螺旋区的芳香族氨基酸含量,提高菌毛导电率[ | 最大电流达130 μA |
过表达甲基转移酶基因phzM,增强菌株胞外电子传递载体-绿脓菌素的合成[ | 最大功率密度达16.67 mW/m2 | |
G.sulfurreducens | 异源表达来自G.metallireducens的pilA基因,增强菌毛导电率[ | pili导电率为277 S/cm |
电活性生物膜的合成生物学改造 | ||
S.oneidensis | 过表达ydeH基因,增强菌株生物膜形成能力[ | 最大功率密度达167.6 mW/m2 |
过表达cyaC基因增加菌株胞内cAMP浓度,提高菌株电流输出密度[ | 最大电流密度达356 mA/m2 | |
G.sulfurreducens | 过表达GSU1501基因促进胞外多糖分泌,促进生物膜形成,提高c型细胞色素丰度[ | Fe(Ⅲ)还原速率达1.75 mmol/(L·d) |
P.aeruginosa | 敲除RpoS基因促进吩嗪类物质合成,提高胞外电子通量[ | 最大电流密度达4.5 μA/cm2 |
过表达irrE基因,强化参与胞内代谢和EET过程[ | 最大功率密度达56.0 mW/m2 | |
构建irrE突变基因文库,增强细胞环境适应性和产电性能[ | 最大功率密度达149.2 mW/m2 |
Tab. 1 Summary of engineering exoelectrogens by synthetic biology
电能细胞 | 合成生物学策略 | 工程结果 |
---|---|---|
底物代谢效率的合成生物学改造 | ||
S.oneidensis | 强化乳酸利用关键因子CRP表达,增强菌株对乳酸的利用能力[ | 最大电流密度约40 mA/m2 |
异源表达视紫红质基因,通过光驱动改变膜电位,增强乳酸的摄取[ | 平均电流密度达0.39 A/m2 | |
异源表达葡萄糖转运基因和葡萄糖激酶基因,增强菌株葡萄糖代谢能力[ | 最大电流输出0.085 mA | |
异源表达木糖转运蛋白、异构酶等构建木糖代谢途径[ | 最大功率密度2.1 mW/m2 | |
异源表达乙酸辅酶A基因(ato1,ato2),强化柠檬酸合酶基因gtlA,构建乙酸代谢路径[ | 最大功率密度8.3 mW/m2 | |
胞内可释放电子池的合成生物学改造 | ||
P.aeruginosa | 过表达NAD+合成酶基因nadE,扩充胞内NAD(H/+)总量[ | 最大功率密度4.0 mW/m2 |
S.oneidensis | 过表达ycel、pncB、nadM、nadD*、nadE*基因,增强NAD+的生物合成[ | 最大功率密度162.8 mW/m2 |
过表达gapA2、pflB、fdh*、mdh基因,强化NADH的合成[ | 最大功率密度105.8 mW/m2 | |
E.coli | 破坏乳酸脱氢酶基因ldhA,释放储存在胞内中间代谢物的电子[ | 最大功率密度3.0 mW/m2 |
过表达非质子泵NADH脱氢酶NDH-2,使更多电子通过氧化呼吸链流入EET途径[ | 输出电流4.7 μA | |
构建C3N代谢通路,获得高效的NAD+代谢途径[ | NAD(H)浓度约6 mmol/L | |
胞外电子传递的合成生物学改造 | ||
E.coli | 通过引入S.oneidensis的MtrCAB色素系统使得E.coli具备胞外电子传递能力[ | Fe(Ⅲ)还原速率达83 μmol/(L·d) |
构建S.oneidensis和E.coli细胞色素融合表达系统[ | 电流密度达到25.32 mA/m2 | |
异源表达PCA合成路径phzA1B1C1D1E1F1G1基因簇,以提高间接胞外电子传递速率[ | 最大功率密度为181.1 mW/m2 | |
S.oneidensis | 过表达内膜细胞色素CymA,加速电子传递至末端周质还原酶[ | 功率达到0.13 mW |
敲除周质色素蛋白napB、fccA、tsdB并过表达细胞色素cctA,降低周质细胞色素复杂性[ | 最高功率密度达到436.5 mW/m2 | |
异源表达来自B.subtilis的核黄素合成基因簇ribADEHC,以提高菌株的胞外电子传递速率[ | 最大功率输出为233.0 mW/m2 | |
强化核黄素表达,结合孔蛋白增强分泌,添加材料rGO,构建三维杂合生物膜系统[ | 最大功率输出达2630 mW/m2 | |
异源构建PCA合成路径和ombB-omaB-omcB-omcS直接电子传递链[ | 最大电流密度达310.2 mA/m2 | |
P.aeruginosa | 通过截断PaPilA C端并增加N端 α-螺旋区的芳香族氨基酸含量,提高菌毛导电率[ | 最大电流达130 μA |
过表达甲基转移酶基因phzM,增强菌株胞外电子传递载体-绿脓菌素的合成[ | 最大功率密度达16.67 mW/m2 | |
G.sulfurreducens | 异源表达来自G.metallireducens的pilA基因,增强菌毛导电率[ | pili导电率为277 S/cm |
电活性生物膜的合成生物学改造 | ||
S.oneidensis | 过表达ydeH基因,增强菌株生物膜形成能力[ | 最大功率密度达167.6 mW/m2 |
过表达cyaC基因增加菌株胞内cAMP浓度,提高菌株电流输出密度[ | 最大电流密度达356 mA/m2 | |
G.sulfurreducens | 过表达GSU1501基因促进胞外多糖分泌,促进生物膜形成,提高c型细胞色素丰度[ | Fe(Ⅲ)还原速率达1.75 mmol/(L·d) |
P.aeruginosa | 敲除RpoS基因促进吩嗪类物质合成,提高胞外电子通量[ | 最大电流密度达4.5 μA/cm2 |
过表达irrE基因,强化参与胞内代谢和EET过程[ | 最大功率密度达56.0 mW/m2 | |
构建irrE突变基因文库,增强细胞环境适应性和产电性能[ | 最大功率密度达149.2 mW/m2 |
电能细胞 | 合成生物学策略 | 工程结果 |
---|---|---|
工程强化噬电细胞电子摄取和转化 | ||
S.oneidensis | 表达光驱动质子泵,强化丁二醇脱氢酶表达并敲除氢化酶基因hyaB、hydA,驱动丁二醇合成[ | 2,3-丁二醇合成量10.06 mmol/L |
E.coli | 异源表达细胞色素蛋白MtrCAB、FccA、CymA碳酸氢盐转运蛋白和碳酸酐酶基因,构建产琥珀酸细胞工厂[ | 琥珀酸产电达30.56 mmol/L |
S.elongatus PCC 7942 | 异源表达色素蛋白OmcS和nif基因,促进N2固定[ | NH3 20 h产率达295.7 μmol/L |
工程噬电细胞代谢路径电合成化学品和生物燃料 | ||
R.eutropha | 敲除聚羟基丁酸合成基因簇,异源表达异丁醇合成路径关键基因alsS、ilvC、ilvD、kivd和yqhD,使代谢流向异丁醇和3-甲基-1-丁醇合成[ | 异丁醇、3-甲基-1-丁醇产量达140 mg/L |
破坏聚羟基丁酸酯合成,并强化异丙醇合成路径,使代谢流向异丙醇合成[ | 异丙醇产量达216 mg/L | |
异源表达核酮糖-1,5-二磷酸羧化酶,强化PHB合成[ | PHB产量达485 mg/L | |
异源过表达的甲羟戊酸途径酶法尼基焦磷酸合酶ERG20、IPP异构酶fni和α-葎草烯合酶(ZSSI),促进α-葎草烯合成[ | α-葎草烯产量达10.8 mg/L | |
表达番茄红素途径基因CrtEBI2,促进番茄红素合成[ | 番茄红素产量达1.73 mg/L | |
S.cerevisiae | 异源表达氧甾醇7α-羟化酶,促进7α-OH-DHEA合成[ | 7α-OH-DHEA产量达288.6 mg/L |
R.palustris | 异源引入丁醇生物合成途径phaABJ、ter、adhE2基因,强化丁醇合成[ | 丁醇产量达0.91 mg/L± 0.07 mg/L |
Tab.2 Summary of engineering electrotrophs by synthetic biology
电能细胞 | 合成生物学策略 | 工程结果 |
---|---|---|
工程强化噬电细胞电子摄取和转化 | ||
S.oneidensis | 表达光驱动质子泵,强化丁二醇脱氢酶表达并敲除氢化酶基因hyaB、hydA,驱动丁二醇合成[ | 2,3-丁二醇合成量10.06 mmol/L |
E.coli | 异源表达细胞色素蛋白MtrCAB、FccA、CymA碳酸氢盐转运蛋白和碳酸酐酶基因,构建产琥珀酸细胞工厂[ | 琥珀酸产电达30.56 mmol/L |
S.elongatus PCC 7942 | 异源表达色素蛋白OmcS和nif基因,促进N2固定[ | NH3 20 h产率达295.7 μmol/L |
工程噬电细胞代谢路径电合成化学品和生物燃料 | ||
R.eutropha | 敲除聚羟基丁酸合成基因簇,异源表达异丁醇合成路径关键基因alsS、ilvC、ilvD、kivd和yqhD,使代谢流向异丁醇和3-甲基-1-丁醇合成[ | 异丁醇、3-甲基-1-丁醇产量达140 mg/L |
破坏聚羟基丁酸酯合成,并强化异丙醇合成路径,使代谢流向异丙醇合成[ | 异丙醇产量达216 mg/L | |
异源表达核酮糖-1,5-二磷酸羧化酶,强化PHB合成[ | PHB产量达485 mg/L | |
异源过表达的甲羟戊酸途径酶法尼基焦磷酸合酶ERG20、IPP异构酶fni和α-葎草烯合酶(ZSSI),促进α-葎草烯合成[ | α-葎草烯产量达10.8 mg/L | |
表达番茄红素途径基因CrtEBI2,促进番茄红素合成[ | 番茄红素产量达1.73 mg/L | |
S.cerevisiae | 异源表达氧甾醇7α-羟化酶,促进7α-OH-DHEA合成[ | 7α-OH-DHEA产量达288.6 mg/L |
R.palustris | 异源引入丁醇生物合成途径phaABJ、ter、adhE2基因,强化丁醇合成[ | 丁醇产量达0.91 mg/L± 0.07 mg/L |
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