Novozymes

About: Novozymes is a company organization based out in Copenhagen, Denmark. It is known for research contribution in the topics: Nucleic acid & Polynucleotide. The organization has 2506 authors who have published 2828 publications receiving 89266 citations. The organization is also known as: Novo Enzymes A/S & Novozymes A/S.

Co-transcription of the celC gene cluster in Clostridium thermocellum

Abstract: Clostridium thermocellum, an anaerobic, thermophilic, and ethanogenic bacterium produces a large cellulase complex termed the cellulosome and many free glycosyl hydrolases. Most cellulase genes scatter around the genome. We mapped the transcripts of the six-gene cluster celC–glyR3–licA–orf4–manB–celT and determined their transcription initiation sites by primer extension. Northern blot showed that celC–glyR3–licA were co-transcribed into a polycistronic messenger with the transcription initiation site at −20 bp. Furthermore, RT-PCR mapping showed that manB and celT, two cellulosomal genes immediately downstream, were co-transcribed into a bicistronic messenger with the initiation site at −233 bp. In contrast, rf4 was transcribed alone with the two initiation sites at −130 and −138 bp, respectively. Finally, quantitative RT-PCR analysis showed that celC, glyR3, and licA were coordinately induced by growing on laminarin, a β-1,3 glucan. Gene expression peaked at the late exponential phase. Taking together with our previous report that GlyR3 binds to the celC promoter in the absence of laminaribiose, a β-1,3 glucose dimer, these results indicate that celC, glyR3, and licA form an operon repressible by GlyR3 and inducible by laminaribiose, signaling the availability of β-1,3 glucan. The celC operon is the first glycosyl hydrolase operon reported in this bacterium.

Biocoatings: challenges to expanding the functionality of waterborne latex coatings by incorporating concentrated living microorganisms

Abstract: Biocoatings concentrate living, nongrowing microbes in nanoporous adhesive polymer films Any microbial activity or trait of interest can be intensified and stabilized in biocoatings These films will dramatically expand the functionality of waterborne coatings Many microbes contain enzyme systems which are unstable when purified Therefore, thin polymer coatings of active microbes are a revolutionary approach to stabilize living cells as industrial or environmental biocatalysts We have demonstrated that some microbes survive polymer film formation embedded in nontoxic adhesive waterborne binders by controlling formulation and drying Biocoatings can be a single layer of randomly oriented microbes or highly structured multilayer films combining monolayers of different types of microbes on solid, porous, or flexible substrates They can be formed by drawdown or ink-jet deposition, convective sedimentation assembly, dielectrophoresis, or coated onto or embedded within papers Controlled drying generates nanoporous microstructure; the pores are filled with a carbohydrate glass which stabilizes the entrapped dehydrated microbes When the coating is rehydrated, the carbohydrates diffuse out generating nanopores The activity of biocoatings can be 100s of g L−1 (coating volume) h−1 stabilized for 100–1000s of hours, and therefore, they represent a new approach to process intensification (PI) using thin liquid film bioreactors A current challenge is that many microbes being engineered as environmental, solar, or carbon recycling biocatalysts do not naturally survive film formation The mechanisms of dehydration damage that occur during biocoating formulation, ambient drying, and during dry storage have begun to be studied Critical to preserving microbe viability are minimizing osmotic stress, toxic monomers, biocides, and utilizing polymer chemistries that generate strong wet adhesion with arrested coalescence (nanoporosity) Therefore, controlling desiccation, drying rate/uniformity, and residual moisture are important Optimization of biocoating activity can be affected at multiple stages—cellular engineering prior to coating (preadaptation), formulation, deposition (film thickness), film formation/drying (generates microstructure), dry storage (minimize metabolic activity), and rehydration Gene induction (activation) leading to enzyme synthesis following rehydration has been demonstrated However, little is known about gene regulation in nongrowing microbes Challenges to optimizing biocoating activity include generating stable film porosity, strong wet adhesion, control of residual water content/form/distribution, and nondestructive measurement of entrapped microbe viability and activity Indirect methods to measure viability include vital staining, enzyme activity, reporter genes, response to light, confocal fluorescent microscopy, and ATP content Microbes containing stress-inducible reporter genes can be used to monitor cell stress during formulation, film formation, and drying Future cellular engineering to optimize biocoatings includes desiccation tolerance, light reactivity (photoefficiency), response to oxidative stress, and cell surface-to-polymer or substrate adhesion Preservation of microbial activity in waterborne coatings could lead to high intensity biocatalysts for environmental cleaning, gaseous carbon recycling, to produce H2 or electricity from microbial fuel cells, delivery of probiotics, or for biosolar energy harvesting

Promoting and Impeding Effects of Lytic Polysaccharide Monooxygenases on Glycoside Hydrolase Activity

Abstract: Lytic polysaccharide monooxygenases (LPMOs) have attracted attention due to their ability to boost cellulolytic enzyme cocktails for application in biorefineries. However, the interplay between LPMOs and individual glycoside hydrolases remains poorly understood. We investigated how the activity of two cellobiohydrolases (Cel7A and Cel6A) and an endoglucanase (Cel7B) from Trichoderma reesei were affected by a C1-oxidizing LPMO from Thielavia terrestris (TtAA9). We quantified products from a mixture of LPMO and glycoside hydrolase and estimated separate contributions of products by each of the enzymes. Hereby, we assessed if an observed synergy reflected a promotion of the activity of hydrolase, LPMO, or both. We consistently found that TtAA9 affected the investigated hydrolases differently. It strongly impeded the turnover of the reducing end cellobiohydrolase, TrCel7A, moderately promoted the turnover of the nonreducing end cellobiohydrolase TrCel6A, and promoted the turnover of the endoglucanase, TrCel7B up to 5-fold. The promoting effect on the endoglucanase increased with hydrolysis extent, indicating that the promoting effect became more important as the recalcitrance of the substrate increased. Experiments with mixtures containing multiple glycoside hydrolases suggested that the LPMO primarily promoted the activity of the endoglucanase, whereas promotion of TrCel6A was secondary.