Production of Chloroplast-Derived Pectin Lyases

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 chloroplast engineering of biofuels enzymes. Our engineers are focused on developing pectin lyases involved in cell wall degradation for textile bioscouring to enhance the water absorption of cotton fibers without causing cellulose damage.

Introduction

Pectin is an abundant cell wall polysaccharide. The enzymes responsible for pectin biosynthesis and modification play a key role in ensuring the specificity and plasticity of cell wall remodeling in different cell types and under different environmental conditions. Among them, pectate lyase is responsible for the eliminative cleavage of pectate, resulting in the production of oligosaccharides with a 4-deoxy-α-D-mann-4-enuronosyl group at its non-reducing end without producing methanol. Large quantities of pectate lyase are required for the paper, food and textile industries. The use of microbial production systems is limited due to expensive fermentation facilities, purification of host cells, and formulations to improve stability. Therefore, there is a great need to explore new production techniques that can eliminate these expensive enzymatic processes. A major recent advance has been the development of chloroplast-derived enzyme cocktails for the production of fermentable sugars from pectin.

Fig. 1. An overview of the synthesis, degradation, and modification of pectin in the plant cell wall. (Kato T, et al., 2018)

Solutions

Chloroplast transformation is a promising technique to obtain high levels of lignocellulolytic enzymes with environmental and biotechnological implications. We have successfully applied chloroplast transformation technology to the development of pectin lyases. Lifeasible is committed to using the PelA gene expressed in tobacco chloroplasts to synthesize pectin lyases.

Our goal is to express cell wall pectin lyases using plastid biotechnology to help you study their expression, stability, and biochemical activity in the chloroplast compartment, and their impact on plant metabolism and growth as well as stable transgene isolation. Our engineered pectin lyases have been widely used to treat residues such as municipal waste, agriculture or papermaking. The strategy for our solution is roughly as follows:

(1) A vector was constructed using the pectate lyase gene (PelA) family 3 from Streptomyces thermocarboxydus.
(2) The pES4 vector containing PelA was used to transform tobacco chloroplasts.
(3) Screening by PCR to detect the presence of PelA in the tobacco chloroplast genome.
(4) Southern blot analysis of the integration, homogeneity and transcriptional stability of PelA protein.
(5) Analyze the effect of pH and temperature on PelA enzyme activity.

Attractive Advantages of Our Solutions

• The pectin lyase protein activity was much higher in transplastid NtpES4 lines than in untransformed plants.
• Providing an economical solution at low cost.
• Pectinases produced in tobacco or lettuce leaves can be tested in the textile or food industry.
• Our dedicated team has performed a genome-wide study of the pectin lyase gene family.

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

References

1. Shin Y, Chane A, Jung M, et al. (2021) Recent Advances in Understanding the Roles of Pectin as an Active Participant in Plant Signaling Networks[J]. Plants. 10(8): 1712.
2. Cao J. (2012) The pectin lyases in Arabidopsis thaliana: evolution, selection and expression profiles. PLoS One. 7(10):e46944.