Efficient carbon dioxide sequestration by using recombinant carbonic anhydrase

Tan, Shih I.; Han, Yin Lung; Yu, You; Chiu, Chen-Yaw; Chang, Yu-Kaung; Ouyang, Shoung; Fan, Kai; Lo, Kuei Ho; Ng, I‐Son

doi:10.1016/j.procbio.2018.08.017

Cited by 39 publications

(27 citation statements)

References 46 publications

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“…In addition, the NaCl also contributed to increase WAU, which is likely to metal effect of SyCA. As the previous result for salinity tolerance of MlCA maintained 82% activity within 6% salinity, 20 SyCA showed higher tolerance, which suggests that SyCA is an outstanding candidate for application in a large-scale CCS. SyCA, but it still remains above 60% of the original activity, while Co +2 , Cu +2 , and Zn +2 will seriously inhibit the activity in both concentrations.…”

Section: Characterization Of Sycasupporting

confidence: 75%

“…The total protein concentration was measured by Bio‐Rad Protein Assay Kits, and the specific CA activity defined by crude enzyme was determined using the Wilbur–Anderson assay . Briefly, 9 mL ice‐cold Tris–HCl (20 mM, pH 8.3) buffer and 0.2 mL enzyme were mixed and transferred to a 20 mL sample bottle, with further incubation at 0°C with stirring.…”

Section: Methodsmentioning

confidence: 99%

“…The total protein concentration was measured by Bio-Rad Protein Assay Kits, and the specific CA activity defined by crude enzyme was determined using the Wilbur-Anderson assay. 20 with the NaCl at a range between 0 and 6% (wt/vol) was substituted.…”

Section: Carbonic Anhydrase Assaymentioning

confidence: 99%

“…In recent years, scientists have attended to apply CA in the CCS process . However, the difficulties are heat resistance and stability of CA.…”

Section: Introductionmentioning

confidence: 99%

“…17 In recent years, scientists have attended to apply CA in the CCS process. [18][19][20] However, the difficulties are heat resistance and stability of CA. Thus, two different strategies to overcome the problem are isolated CA from species that survive in extreme environments, that is, hot springs or anaerobic conditions, 21 or to engineer protein by genetic approach.…”

mentioning

confidence: 99%

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ARduino‐pH Tracker and screening platform for characterization of recombinant carbonic anhydrase in Escherichia coli

Hsu

Tan

Chiu

et al. 2019

Biotechnology Progress

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Carbonic anhydrase (CA, EC 4.2.1.1) is an ancient enzyme with zinc ion as its active site, which catalyzes the chemical reaction of carbon dioxide (CO2) to react with water and form bicarbonate ions. Due to its high catalytic efficiency on CO2 assimilation, CA is expected to use for carbon sequestration in industry. However, the protein expression level, thermostability and high‐throughput screening of an active CA are still with difficulty. In this study, the CA from Sulfurihydrogenibium yellowstonense (denoted as SyCA) was selected for overexpressed in Escherichia coli by different pET vectors. The enzymatic properties including thermo‐stability, pH tolerance, effect of metal ion, and kinetic parameters were characterized through a novel ARduino‐pH Tracker (ART) for monitoring online effectively. The SyCA is thermophilic and acidophilic as it maintains 100% activity at 50°C, while the residual activity is 34.8% after heating at 80°C for 150 min and the optimal pH is 3–5. The kinetic analysis by ART system showed that the k cat/K m of free enzyme was 4.4‐folds that that of whole cell. On the other hand, the screening platforms as Wilbur–Anderson unit, phenol red indicator and size of colony forming unit have been established to explore CA with higher activity. The high‐throughput screening platform is support in direct evolution of CA and further used in the industry.

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Section: Characterization Of Sycasupporting

confidence: 75%

Section: Methodsmentioning

confidence: 99%

Section: Carbonic Anhydrase Assaymentioning

confidence: 99%

“…In recent years, scientists have attended to apply CA in the CCS process . However, the difficulties are heat resistance and stability of CA.…”

Section: Introductionmentioning

confidence: 99%

mentioning

confidence: 99%

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ARduino‐pH Tracker and screening platform for characterization of recombinant carbonic anhydrase in Escherichia coli

Hsu

Tan

Chiu

et al. 2019

Biotechnology Progress

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CRISPRi‐mediated programming essential gene can as a Direct Enzymatic Performance Evaluation & Determination (DEPEND) system

Tan

2020

Biotech & Bioengineering

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Harnessing enzyme expression for production of target chemicals is a critical and multifarious process, where screening of different genes by inspection of enzymatic activity plays an imperative role. Here, we conceived an idea to improve the time‐consuming and labor‐intensive process of enzyme screening. Controlling cell growth was achieved by the Cluster Regularly Interspaced Short Palindromic Repeat (CRISPRi) system with different single guide RNA targeting the essential gene can (CRISPRi::CA) that encodes a carbonic anhydrase for CO2 uptake. CRISPRi::CA comprises a whole‐cell biosensor to monitor CO2 concentration, ranging from 1% to 5%. On the basis of CRISPRi::CA, an effective and simple Direct Enzymatic Performance Evaluation & Determination (DEPEND) system was developed by a single step of plasmid transformation for targeted enzymes. As a result, the activity of different carbonic anhydrases corresponded to the colony‐forming units. Furthermore, the enzymatic performance of 5‐aminolevulinic acid synthetase (ALAS), which converts glycine and succinate‐CoA to release a molecule of CO2, has also been distinguished, and the effect of the chaperone GroELS on ALAS enzyme folding was successfully identified in the DEPEND system. We provide a highly feasible, time‐saving, and flexible technology for the screening and inspection of high‐performance enzymes, which may accelerate protein engineering in the future.

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