Insights into Cell-Free Conversion of CO₂ to Chemicals by a Multienzyme Cascade Reaction

TY - JOUR

T1 - Insights into Cell-Free Conversion of CO2 to Chemicals by a Multienzyme Cascade Reaction

AU - Singh, Raushan Kumar

AU - Singh, Ranjitha

AU - Sivakumar, Dakshinamurthy

AU - Kondaveeti, Sanath

AU - Kim, Taedoo

AU - Li, Jinglin

AU - Sung, Bong Hyun

AU - Cho, Byung Kwan

AU - Kim, Dong Rip

AU - Kim, Sun Chang

AU - Kalia, Vipin C.

AU - Zhang, Yi Heng P.Job

AU - Zhao, Huimin

AU - Kang, Yun Chan

AU - Lee, Jung Kul

PY - 2018/12/7

Y1 - 2018/12/7

N2 - Multienzymatic cascade reactions have garnered the attention of many researchers as an approach for converting CO2 into methanol. The cascade reaction used in this study includes the following enzymes: a formate dehydrogenase (ClFDH), a formaldehyde dehydrogenase (BmFaldDH), and an alcohol dehydrogenase (YADH) from Clostridium ljungdahlii, Burkholderia multivorans, and Saccharomyces cerevisiae, respectively. Because this cascade reaction requires NADH as a cofactor, phosphite dehydrogenase (PTDH) was employed to regenerate the cofactor. The multienzymatic cascade reaction, along with PTDH, yielded 3.28 mM methanol. The key to the success of this cascade reaction was a novel formaldehyde dehydrogenase, BmFaldDH, the enzyme catalyzing the reduction of formate to formaldehyde. The methanol yield was further improved by incorporation of 1-ethyl-3-methylimidazolium acetate (EMIM-Ac), resulting in 7.86 mM of methanol. A 500-fold increase in total turnover number was observed for the ClFDH-BmFaldDH-YADH cascade system compared to the Candida boidinii FDH-Pseudomonas putida FaldDH-YADH system. We provided detailed insights into the enzymatic reduction of CO2 by determining the thermodynamic parameters (Kd and ΔG) using isothermal titration calorimetry. Furthermore, we demonstrated a novel time-dependent formaldehyde production from CO2. Our results will aid in the understanding and development of a robust multienzyme catalyzed cascade reaction for the reduction of CO2 to value-added chemicals.

AB - Multienzymatic cascade reactions have garnered the attention of many researchers as an approach for converting CO2 into methanol. The cascade reaction used in this study includes the following enzymes: a formate dehydrogenase (ClFDH), a formaldehyde dehydrogenase (BmFaldDH), and an alcohol dehydrogenase (YADH) from Clostridium ljungdahlii, Burkholderia multivorans, and Saccharomyces cerevisiae, respectively. Because this cascade reaction requires NADH as a cofactor, phosphite dehydrogenase (PTDH) was employed to regenerate the cofactor. The multienzymatic cascade reaction, along with PTDH, yielded 3.28 mM methanol. The key to the success of this cascade reaction was a novel formaldehyde dehydrogenase, BmFaldDH, the enzyme catalyzing the reduction of formate to formaldehyde. The methanol yield was further improved by incorporation of 1-ethyl-3-methylimidazolium acetate (EMIM-Ac), resulting in 7.86 mM of methanol. A 500-fold increase in total turnover number was observed for the ClFDH-BmFaldDH-YADH cascade system compared to the Candida boidinii FDH-Pseudomonas putida FaldDH-YADH system. We provided detailed insights into the enzymatic reduction of CO2 by determining the thermodynamic parameters (Kd and ΔG) using isothermal titration calorimetry. Furthermore, we demonstrated a novel time-dependent formaldehyde production from CO2. Our results will aid in the understanding and development of a robust multienzyme catalyzed cascade reaction for the reduction of CO2 to value-added chemicals.

KW - cascade reaction

KW - CO

KW - FaldDH

KW - FDH

KW - formaldehyde

KW - methanol

KW - multienzyme

UR - http://www.scopus.com/inward/record.url?scp=85058292332&partnerID=8YFLogxK

U2 - 10.1021/acscatal.8b02646

DO - 10.1021/acscatal.8b02646

M3 - Journal article

AN - SCOPUS:85058292332

SN - 2155-5435

VL - 8

SP - 11085

EP - 11093

JO - ACS Catalysis

JF - ACS Catalysis

IS - 12

ER -