Enzyme mediated transformation of CO2 into calcium carbonate using purified microbial carbonic anhydrase

Tanvi Sharma; Abhishek Sharma; Chang Lei Xia; Su Shiung Lam; Azmat Ali Khan; Sonam Tripathi; Raj Kumar; Vijai Kumar Gupta; Ashok Kumar Nadda

doi:10.1016/j.envres.2022.113538

Enzyme mediated transformation of CO₂ into calcium carbonate using purified microbial carbonic anhydrase

Environ Res. 2022 Sep;212(Pt D):113538. doi: 10.1016/j.envres.2022.113538. Epub 2022 May 28.

Authors

Tanvi Sharma¹, Abhishek Sharma², Chang Lei Xia³, Su Shiung Lam⁴, Azmat Ali Khan⁵, Sonam Tripathi⁶, Raj Kumar¹, Vijai Kumar Gupta⁷, Ashok Kumar Nadda⁸

Affiliations

¹ Department of Biotechnology and Bioinformatics, Jaypee University of Information Technology, Waknaghat, Solan, Himachal Pradesh, 173 234, India.
² Department of Biotechnology, Himachal Pradesh University, Summer Hill, Shimla, 171 005, India.
³ Co-Innovation Center of Efficient Processing and Utilization of Forestry Resources, College of Materials Science and Engineering, Nanjing Forestry University, Nanjing, Jiangsu, 210037, China.
⁴ Higher Institution Centre of Excellence (HICoE), Institute of Tropical Aquaculture and Fisheries (AKUATROP), Universiti Malaysia Terengganu, 21030 Kuala Nerus, Terengganu, Malaysia.
⁵ Pharmaceutical Biotechnological Laboratory, Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, Riyadh, 11451, Saudi Arabia.
⁶ Department of Environmental Microbiology, School for Environmental Sciences, Babasaheb Bhimrao Ambedkar Central University, Lucknow, Uttar Pradesh, 226025, India.
⁷ Biorefining and Advanced Materials Research Center, Scotland's Rural College (SRUC), Kings Buildings, West Mains Road, Edinburgh, EH9 3JG, UK.
⁸ Department of Biotechnology and Bioinformatics, Jaypee University of Information Technology, Waknaghat, Solan, Himachal Pradesh, 173 234, India. Electronic address: ashok.nadda@juit.ac.in.

PMID: 35640707
DOI: 10.1016/j.envres.2022.113538

Abstract

In this study, a bacterial carbonic anhydrase (CA) was purified from Corynebacterium flavescens for the CO₂ conversion into CaCO₃. The synthesized CaCO₃ can be utilized in the papermaking industry as filler material, construction material and in steel industry. Herein, the CA was purified by using a Sephadex G-100 column chromatography having 29.00 kDa molecular mass in SDS-PAGE analysis. The purified CA showed an optimal temperature of 35 °C and pH 7.5. In addition, a kinetic study of CA using p-NPA as substrate showed V_max (166.66 μmoL/mL/min), K_m (5.12 mM), and K_cat (80.56 sec^-1) using Lineweaver Burk plot. The major inhibitors of CA activity were Na²⁺, K⁺, Mn²⁺, and Al³⁺, whereas Zn²⁺ and Fe²⁺ slightly enhanced it. The purified CA showed a good efficacy to convert the CO₂ into CaCO₃ with a total conversion rate of 65.05 mg CaCO₃/mg of protein. In silico analysis suggested that the purified CA has conserved Zn²⁺ coordinating residues such as His 111, His 113, and His 130 in the active site center. Further analysis of the CO₂ binding site showed conserved residues such as Val 132, Val 142, Leu 196, Thr 197, and Val 205. However, a substitution has been observed where Trp 208 of its closest structural homolog T. ammonificans CA is replaced with Arg 207 of C. flavescens. The presence of a hydrophilic mutation in the CO₂ binding hydrophobic region is a further subject of investigation.

Keywords: CO(2) conversion; CaCO(3); Carbonic anhydrase; In silico; Purification.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Calcium Carbonate
Carbon Dioxide / chemistry
Carbonic Anhydrases* / chemistry
Carbonic Anhydrases* / genetics
Carbonic Anhydrases* / metabolism
Electrophoresis, Polyacrylamide Gel
Temperature

Substances

Carbon Dioxide
Carbonic Anhydrases
Calcium Carbonate