Production of Chloroplast-Derived β-Glucosidases

Plant chloroplast genetic engineering can utilize transgenic crops to express different enzymes from bacteria or fungi for the production of feedstocks for biofuels. Lifeasible has successfully developed a variety of reliable and economical solutions for enzymatic chloroplast engineering for biofuels. Our engineers focus on developing β-glucosidases for the biofuel industry, facilitating the biotransformation of cellulosic biomass.

Introduction

β-Glucosidase (BGL) is a dual-characterized enzyme with both glycosidic bond synthesis and degradation, which makes it an enzyme with great industrial potential. Beta-glucosidases can convert cellobiose and cellooligosaccharides into glucose monomers, which are then used to produce biofuels through a fermentation process. Due to low yields, lack of potential BGL-producing microorganisms, and low activity, the overall conversion of cellulose to sugars is low, and large-scale production of biofuels to replace fossil fuels is not feasible. Various beta-glucosidase genes have been overexpressed in plants. In addition, the ability of chloroplasts to express multiple genes provides the potential to express β-glucosidase.

Fig. 1. Hydrolysis of cellulose by the synergistic action of cellulases. (Srivastava N, et al., 2019)

Solutions

Chloroplast genetic engineering has allowed the design, integration and expression of a variety of genes to produce high levels of agro-industrial variety of proteins at low cost. We have successfully applied chloroplast transformation technology to the development of β-glucosidase. Lifeasible is committed to the synthesis of β-glucosidase using the bgl1 gene expressed in tobacco chloroplasts. Faced with the challenge of β-glucosidase production of biofuels, we offer the following strategies to provide you with an economical solution for the production of chloroplast-derived β-glucosidase:

• Selecting effective microbial strains to reduce production costs, such as Penicillium.
• A suitable substrate for the production of β-glucosidase was selected by combining carbon sources, nitrogen sources and inducers.
• Increasing the quantity and efficiency of beta-glucosidase.
• Inhibiting β-glucosidase by glucose, thereby blocking substrate binding to the active site of β-glucosidase.
• β-glucosidase was reused by immobilization.
• Enhancing β-glucosidase activity and stability over a wide range of pH and temperature.

Our customers have successfully used chloroplast-derived β-glucosidases for cell wall degradation for different industrial uses, including residue treatment of municipal waste, agriculture or papermaking. The strategy for our solution is roughly as follows:

(1) A vector was constructed using b-glucosidase (bgl1 gene) from Aspergillus niger.
(2) Transform the vector into tobacco chloroplasts.
(3) PCR screening to detect the presence of bgl1 gene in tobacco chloroplast genome.
(4) Southern blot analysis of bgl1 protein integration, homogeneity and transcriptional stability.
(5) β-glucosidase activity was estimated by quantification of pnitrophenol released from p-nitrophenyl-β-D-glucopyranoside (pNPG).

Attractive Advantages of Our Solutions

• Chloroplast-derived β-glucosidase has high yield and high activity.
• Expression of β-glucosidase in different organisms can be achieved.
• Chloroplast-derived beta-glucosidases can increase plant hormone levels, increase biomass, increase trichomes, and provide protection against aphids or whiteflies.
• The bgl1 gene can be integrated and expressed in tobacco chloroplasts with high production of cellulohydrolase.
• Large-scale and sustainable production of β-glucosidase using plant systems.

Lifeasible is committed to providing customers around the world with fully customized chloroplast engineered solutions for β-glucosidases production. Please contact us to discuss further details to ensure your next success.

References

1. Srivastava N, Rathour R, Jha S, et al. (2019) Microbial Beta Glucosidase Enzymes: Recent Advances in Biomass Conversation for Biofuels Application. Biomolecules. 9(6): 220.
2. Espinoza-Sánchez E A, Torres-Castillo J A, Rascón-Cruz Q, et al. (2016) Production and characterization of fungal β-glucosidase and bacterial cellulases by tobacco chloroplast transformation[J]. Plant Biotechnology Reports. 10(2): 61-73.