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Role of Cel5H protein surface amino acids in binding with clay minerals and measurements of its forces

ABSTRACT: Our previous study on the binding activity between Cel5H and clay minerals showed highest binding efficiency among other cellulase enzymes cloned. Here, based on previous studies, we hypothesized that the positive amino acids on the surface of Cel5H protein may play an important role in binding to clay surfaces. To examine this, protein sequences of Bacillus licheniformis Cel5H (BlCel5H) and Paenibacillus polymyxa Cel5A (PpCel5A) were analyzed and then selected amino acids were mutated. These mutated proteins were investigated for binding activity and force measurement via atomic force microscopy (AFM). A total of seven amino acids which are only present in BlCel5H but not in PpCel5A were selected for mutational studies and the positive residues which are present in both were omitted. Of the seven selected surface lysine residues, only three mutants K196A(M2), K54A(M3) and K157T(M4) showed 12%, 7% and 8% less clay mineral binding ability, respectively compared with wild-type. The probable reason why other mutants did not show altered binding efficiency might be due to relative location of amino acids on the protein surface. Meanwhile, measurement of adhesion forces on mica sheets showed a well-defined maximum at 69 ± 19 pN for wild-type, 58 ± 19 pN for M2, 53 ± 19 pN for M3, and 49 ± 19 pN for M4 proteins. Hence, our results demonstrated that relative location of surface amino acids of Cel5H protein especially positive charged amino acids are important in the process of clay mineral-protein binding interaction through electrostatic exchange of charges.

Supplementary Information

The online version contains supplementary material available at 10.1186/s42649-021-00066-7.

SUBMITTER: Math R

PROVIDER: S-EPMC8586110 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Simple Summary Acheta domesticus (house cricket) flour is one of the most promising and sustainable sources of nutrients. It is rich in protein, minerals, fatty acids, amino acids, and vitamins and it has a low impact on the consumption of natural environmental resources. On the other side, fermentation with lactic acid bacteria represents a technological tool that can improve further the nutritional quality of flours. Therefore, the aim of the present research was to study the adaptability of the Lactobacillus plantarum strain on insect flour fermentation. Fatty acids, amino acids, minerals, and aroma volatile compounds were analyzed during 48 h of fermentation. Fermentation improved the nutritional quantity of the bioactive compounds, mainly after 24 h of fermentation, where they reached higher extended values. Overall, our findings indicate that insect flour is able to support the growth and development of the Lactobacillus plantarum strain, leading to an enriched insect flour sourdough that could be further used in the manufacturing of new products. Abstract Acheta domesticus (L.1758) has been recently accepted by the European Union as a novel food, being the third insect that has been approved for human consumption. Nowadays, researchers’ attention is focused on exploiting new protein sustainable sources, and, therefore, insect flour has gained more and more interest. Organic acids, fatty acids, amino acids, aroma volatile compounds, and minerals were analyzed through HPLC-RID (High-performance liquid chromatography), GC-MS (Gas chromatography-mass spectrometry), LC-MS (Liquid chromatography–mass spectrometry), ITEX/GC-MS and AAS (Atomic Absorption Spectrophotometry), respectively. Fermentation of the insect flour with Lactobacillus plantarum ATCC 8014 strain (Lp) leads to an increase in organic acids such as lactic, acetic, and oxalic, whilst citric acid decreases its value. SFA (saturated fatty acids) and MUFA (monosaturated fatty acids) groups were positively influenced by Lp fermentation; meanwhile, PUFA (polysaturated fatty acids) decreased during fermentation. A positive trend was observed for amino acids, aroma volatile content, and minerals enhancement during insect sourdough fermentation, mainly at 24 h of fermentation. Acheta domesticus (A. domesticus) sourdough fermentation represents a new tool that needs to be further exploited aiming to improve the nutritional qualities of the final products.

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Role of Cel5H protein surface amino acids in binding with clay minerals and measurements of its forces

Supplementary Information

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