Rational Design of K173A Substitution Enhances Thermostability Coupled with Catalytic Activity of &lt;b&gt;&lt;i&gt;Enterobacter&lt;/i&gt;&lt;/b&gt; sp. Bn12 Lipase

Farrokh, Parisa; Yakhchali, Bagher; Karkhane, Ali Asghar

doi:10.1159/000365890

Cited by 6 publications

(11 citation statements)

References 40 publications

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“…Generally, in the case of surfactants, the activity of the Q177A lipase increased in the presence of nonionic surfactants (1% w/v) and decreased in the ionic ones (Table ), similar to the native and K173A lipases . The K173A/Q177A lipase also showed the same activity pattern, whereas Triton X‐100 had no significant effect on its activity.…”

Section: Resultsmentioning

confidence: 68%

“…The optimum activity of the Q177A and K173A/Q177A mutants was observed at pH 8.0 (data not shown) and it was similar to that of native and K173A lipases . pH stability analysis exhibited that native, K173A, and Q177A lipases retained more activity than the double‐mutant lipase after 1‐H incubation in alkaline buffers (pH 9.0–11) (Fig.…”

Section: Resultsmentioning

confidence: 71%

“…The plasmids pYKF and pYKF‐K173A were used as templates for construction of single‐mutant (Q177A) and double‐mutant (K173A/Q177A) lipases through overlap extension PCR, respectively . Construction of the mutated lipases was performed with primer sets of YKF‐1/Q177A‐1 and YKF‐2/Q177A‐2 (Table ) in two rounds of PCR, and the full‐length genes were amplified with YKF‐1 and YKF‐2 primers.…”

Section: Methodsmentioning

confidence: 99%

“…The K173A/Q177A lipase variant retained its activity the same as the native except for the acetone, which decreased the enzyme activity (74.4%) ( Table 4). The activity of the lipases in the polar organic solvents shows that substitution of the nonpolar Ala with polar Lys/Gln in the surface of the variants does not necessarily have an adverse impact [17]. It seems that positive effect of organic solvents on the Q177A lipase could be due to enzyme structural changes [41].…”

Section: Effect Of Organic Solvents Surfactants Metal Ions and Edtmentioning

confidence: 99%

“…Lipase protein engineering for upgrading enzyme properties has been done since the 1980s . Both directed evolution and rational design were successfully applied to improve lipase characteristics in the previous investigations . For example, thermostability and catalytic activity of Bacillus licheniformis lipase were increased through error‐prone PCR and the mutant lipase displayed 13.5‐fold higher thermostability than the wild‐type lipase .…”

Section: Introductionmentioning

confidence: 99%

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Role of Q177A and K173A/Q177A substitutions in thermostability and activity of the ELBn12 lipase

Farrokh

Yakhchali

Karkhane

2017

Biotech and App Biochem

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Thermostable lipases have many applications in detergent industries and in organic synthesis. There are many ways to improve thermal stability of enzymes, for example, higher hydrophobicity, greater structural packing, higher content of the charged residues, and lower thermolabile ones. In this study, thermolabile Gln (sensitive to higher temperatures) was substituted with Ala in native ELBn12 and mutated K173A lipases to examine its effect on thermal stability and activity of the lipases. Single (Q177A) and double mutants (K173A/Q177A) were expressed in Escherichia coli pLysS. The Q177A variant increased both activity and thermostability of the lipase, whereas K173A/Q177A had a negative effect on the lipase activity and only had better thermal stability than the native at 50 °C. pH stability of the double mutant between 9.0 and 11 was also lower than the other variants. Stability analysis in the presence of chemicals showed that Q177A mutant had better activity with 50% (v/v) organic solvents. On the other hand, K173A lipase showed increased activity with 1% (w/v) nonionic surfactant, and finally K173A/Q177A had better stability with 10 mM metal ions compared to the native lipase.

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Section: Resultsmentioning

confidence: 68%

Section: Resultsmentioning

confidence: 71%

Section: Methodsmentioning

confidence: 99%

Section: Effect Of Organic Solvents Surfactants Metal Ions and Edtmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Role of Q177A and K173A/Q177A substitutions in thermostability and activity of the ELBn12 lipase

Farrokh

Yakhchali

Karkhane

2017

Biotech and App Biochem

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Cyanobacteria as an eco‐friendly resource for biofuel production: A critical review

Farrokh

Sheikhpour

Kasaeian³

et al. 2019

Biotechnology Progress

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Cyanobacteria are photosynthetic microorganisms which can be found in various environmental habitats. These photosynthetic bacteria are considered as promising feedstock for the production of the third‐ and the fourth‐generation biofuels. The main subject of this review is highlighting the significant aspects of the biofuel production from cyanobacteria. The most recent investigations about the extraction or separation of the bio‐oil from cyanobacteria are also adduced in the present review. Moreover, the genetic engineering of cyanobacteria for improving biofuel production and the impact of bioinformatics studies on the designing better‐engineered strains are mentioned. The large‐scale biofuel production is challenging, so the economic considerations to provide inexpensive biofuels are also cited. It seems that the future of biofuels is strongly dependent to the following items; understanding the metabolic pathways of the cyanobacterial species, progression in the construction of the engineered cyanobacteria, and inexpensive large‐scale cultivation of them.

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Thermostable lipases and their dynamics of improved enzymatic properties

Hamdan

Maiangwa

Ali

et al. 2021

Appl Microbiol Biotechnol

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Thermal stability is one of the most desirable characteristics in the search for novel lipases. The search for thermophilic microorganisms for synthesising functional enzyme biocatalysts with the ability to withstand high temperature, and capacity to maintain their native state in extreme conditions opens up new opportunities for their biotechnological applications. Thermophilic organisms are one of the most favoured organisms, whose distinctive characteristics are extremely related to their cellular constituent particularly biologically active proteins. Modifications on the enzyme structure are critical in optimizing the stability of enzyme to thermophilic conditions. Thermostable lipases are one of the most favourable enzymes used in food industries, pharmaceutical field, and actively been studied as potential biocatalyst in biodiesel production and other biotechnology application. Particularly, there is a trade-off between the use of enzymes in high concentration of organic solvents and product generation. Enhancement of the enzyme stability needs to be achieved for them to maintain their enzymatic activity regardless the environment. Various approaches on protein modification applied since decades ago conveyed a better understanding on how to improve the enzymatic properties in thermophilic bacteria. In fact, preliminary approach using advanced computational analysis is practically conducted before any modification is being performed experimentally. Apart from that, isolation of novel extremozymes from various microorganisms are offering great frontier in explaining the crucial native interaction within the molecules which could help in protein engineering. In this review, the thermostability prospect of lipases and the utility of protein engineering insights into achieving functional industrial usefulness at their high temperature habitat are highlighted. Similarly, the underlying thermodynamic and structural basis that defines the forces that stabilize these thermostable lipase is discussed.

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Rational Design of K173A Substitution Enhances Thermostability Coupled with Catalytic Activity of <b><i>Enterobacter</i></b> sp. Bn12 Lipase

Cited by 6 publications

References 40 publications

Role of Q177A and K173A/Q177A substitutions in thermostability and activity of the ELBn12 lipase

Role of Q177A and K173A/Q177A substitutions in thermostability and activity of the ELBn12 lipase

Cyanobacteria as an eco‐friendly resource for biofuel production: A critical review

Thermostable lipases and their dynamics of improved enzymatic properties

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