Validation of leaf enzymes in the detergent and textile industries: launching of a new platform technology

Abstract

Summary Chemical catalysts are being replaced by biocatalysts in almost all industrial applications due to environmental concerns, thereby increasing their demand. Enzymes used in current industries are produced in microbial systems or plant seeds. We report here five newly launched leaf‐enzyme products and their validation with 15 commercial microbial‐enzyme products, for detergent or textile industries. Enzymes expressed in chloroplasts are functional at broad pH /temperature ranges as crude‐leaf extracts, while most purified commercial enzymes showed significant loss at alkaline pH or higher temperature, required for broad range commercial applications. In contrast to commercial liquid enzymes requiring cold storage/transportation, chloroplast enzymes as a leaf powder can be stored up to 16 months at ambient temperature without loss of enzyme activity. Chloroplast‐derived enzymes are stable in crude‐leaf extracts without addition of protease inhibitors. Leaf lipase/mannanase crude extracts removed chocolate or mustard oil stains effectively at both low and high temperatures. Moreover, leaf lipase or mannanase crude‐extracts removed stain more efficiently at 70 °C than commercial microbial enzymes (<10% activity). Endoglucanase and exoglucanase in crude leaf extracts removed dye efficiently from denim surface and depilled knitted fabric by removal of horizontal fibre strands. Due to an increased demand for enzymes in the food industry, marker‐free lettuce plants expressing lipase or cellobiohydrolase were created for the first time and site‐specific transgene integration/homoplasmy was confirmed by Southern blots. Thus, leaf‐production platform offers a novel low‐cost approach by the elimination of fermentation, purification, concentration, formulation and cold‐chain storage/transportation. This is the first report of commercially launched protein products made in leaves and validated with current commercial products.

Role of Small Intestine and Gut Microbiome in Plant-Based Oral Tolerance for Hemophilia

“…Instead, the enzymatic activities are contributed by other types of bacteria that are present but infrequent in the small intestine, such as Burkholderiales or Flavobacteriales. Therefore, Daniell et al have recently generated plants expressing cell wall degrading enzymes (24,25). Importantly, oral delivery of pectinase, endoglucanase, exoglucanase, and mannanase before feeding plants cells expressing therapeutic proteins almost doubled drug levels in plasma (46).…”

“…Commercial scale production in cGMP hydroponic facility has been demonstrated for several human blood proteins (16,20,24). Most importantly, methods have been developed to remove antibiotic resistance genes from chloroplast genomes of edible plant cells producing enzymes or biopharmaceuticals (20,24,25). Plant cell wall protects antigens from acid and enzymes in the stomach because they do not cleave Beta1-4, 1-6 linkages in plant cell wall polymers (17,26).…”

From δ-aminolevulinic acid to chlorophylls and every step in between: in memory of Constantin (Tino) A. Rebeiz, 1936–2019

“…This research was continued when I became an independent faculty member at Washington State University, where isolated etio-chloroplasts were first used to express "foreign" genes, resulting in a breakthrough publication (Daniell and McFadden 1987), and we obtained the first and the oldest patent in the field of chloroplast genetic engineering (US patent # 5,693,507). Today, the chloroplast biotechnology field has grown to such an extent that there are regular Gordon Research Conferences on this topic, and research in this area has produced commercial (Daniell et al 2019;Kumari et al 2019) and low-cost drug development (Daniell et al 2016).…”

From conception to COVID‐19: an arduous journey of tribulations of racism and triumphs

“…After bioscouring with pectinase, they readily absorbed water in a few milliseconds, meeting industry requirements. Similarly, four newly launched leaf-enzyme products (endoglucanase, exoglucanase, lipase, mannanase) were also compared and validated with 15 commercial microbial-enzyme products for the detergent and textile industries (Kumari et al, 2019). Leaflipase/mannanase crude extracts removed chocolate or mustard oil stains effectively at both low and high temperatures.…”

“…Learning valuable lessons from the failure of Chlorogen and unwilling to give up, I persuaded investors of the biotechnology start‐up PhylloZyme – who also have ownership in the textile and microbial‐enzyme business – to develop products that required minimal regulatory approv al . Most current genetically modified plant commercial products on the market are derived from seeds, and therefore, it was quite thrilling to launch a first leaf‐made protein product for commercial use developed by PhylloZyme in the summer of 2018 (Daniell et al ., 2019c; Kumari et al ., 2019). Dry leaf‐pectinases were validated with eight liquid commercial microbial‐enzyme products for textile or juice industry applications.…”