Seafood Processing Chitin Waste for Electricity Generation in a Microbial Fuel Cell Using Halotolerant Catalyst Oceanisphaera arctica YHY1

In this study, a newly isolated halotolerant strain Oceanisphaera arctica YHY1, capable of hydrolyzing seafood processing waste chitin biomass, is reported. Microbial fuel cells fed with 1% chitin and 40 g L−1 as the optimum salt concentration demonstrated stable electricity generation until 216 h (0.228 mA/cm2). N-acetyl-D-glucosamine (GlcNAc) was the main by-product in the chitin degradation, reaching a maximum concentration of 192.01 mg g−1 chitin at 120 h, whereas lactate, acetate, propionate, and butyrate were the major metabolites detected in the chitin degradation. O. arctica YHY1 utilized the produced GlcNAc, lactate, acetate, and propionate as the electron donors to generate the electric current. Cyclic voltammetry (CV) investigation revealed the participation of outer membrane-bound cytochromes, with extracellular redox mediators partly involved in the electron transfer mechanism. Furthermore, the changes in structural and functional groups in chitin after degradation were analyzed using FTIR and XRD. Therefore, the ability of O. arctica YHY1 to utilize waste chitin biomass under high salinities can be explored to treat seafood processing brine or high salt wastewater containing chitin with concurrent electricity generation.

Physiological characterization of chitin synthase A responsible for the biosynthesis of cuticle chitin in Culex pipiens pallens (Diptera: Culicidae)

Background The pathogens transmitted by mosquitoes to humans and animals cause several emerging and resurgent infectious diseases. Increasing insecticide resistance requires rational action to control the target vector population. Chitin is indispensable for insect growth and development and absent from vertebrates and higher plants. Chitin synthase A (CHSA) is a crucial enzyme in chitin synthesis; therefore, identifying and characterizing how CHSA determines chitin content may contribute to the development of novel vector control strategies. Results The injection of small interfering RNA targeting CHSA (siCHSA) to knockdown CHSA transcripts in larval, pupal and adult stages of Culex pipiens pallens resulted in the appearance of different lethal phenotypes. When larval and pupal stages were injected with siCHSA, CHSA knockdown prevented larval molting, pupation and adult eclosion, and affected the production of chitin and chitin degradation, which resulted in an ecdysis defect phenotype of mosquitoes. When siCHSA was injected into mosquitoes in the adult stage, CHSA knockdown also affected the laminar organization of the mesoderm and the formation of pseudo-orthogonal patterns of the large fibers of the endoderm. Conclusion We provide a systematic and comprehensive description of the effects of CHSA on morphogenesis and metamorphosis. The results show that CHSA not only affects chitin synthesis during molting, but also might be involved in chitin degradation. Our results further show that CHSA is important for the structural integrity of the adult mosquito cuticle. Graphic abstract

Chitin Synthesis and Degradation in Crustaceans: A Genomic View and Application

Chitin is among the most important components of the crustacean cuticular exoskeleton and intestinal peritrophic matrix. With the progress of genomics and sequencing technology, a large number of gene sequences related to chitin metabolism have been deposited in the GenBank database in recent years. Here, we summarized the genes and pathways associated with the biosynthesis and degradation of chitins in crustaceans based on genomic analyses. We found that chitin biosynthesis genes typically occur in single or two copies, whereas chitin degradation genes are all multiple copies. Moreover, the chitinase genes are significantly expanded in most crustacean genomes. The gene structure and expression pattern of these genes are similar to those of insects, albeit with some specific characteristics. Additionally, the potential applications of the chitin metabolism genes in molting regulation and immune defense, as well as industrial chitin degradation and production, are also summarized in this review.

Functional metagenomics reveals differential chitin degradation and utilization features across free-living and host-associated marine microbiomes

BackgroundChitin ranks as the most abundant polysaccharide in the oceans yet knowledge of shifts in structure and diversity of chitin-degrading communities across marine niches is scarce. Here, we integrate cultivation-dependent and -independent approaches to shed light on the chitin processing potential within the microbiomes of marine sponges, octocorals, sediments, and seawater.ResultsWe found that cultivatable host-associated bacteria in the generaAquimarina,Enterovibrio,Microbulbifer,Pseudoalteromonas,Shewanella, andVibriowere able to degrade colloidal chitin in vitro. Congruent with enzymatic activity bioassays, genome-wide inspection of cultivated symbionts revealed thatVibrioandAquimarinaspecies, particularly, possess several endo- and exo-chitinase-encoding genes underlying their ability to cleave the large chitin polymer into oligomers and dimers. Conversely,Alphaproteobacteriaspecies were found to specialize in the utilization of the chitin monomer N-acetylglucosamine more often. Phylogenetic assessments uncovered a high degree of within-genome diversification of multiple, full-length endo-chitinase genes forAquimarinaandVibriostrains, suggestive of a versatile chitin catabolism aptitude. We then analyzed the abundance distributions of chitin metabolism-related genes across 30 Illumina-sequenced microbial metagenomes and found that the endosymbiotic consortium ofSpongia officinalisis enriched in polysaccharide deacetylases, suggesting the ability of the marine sponge microbiome to convert chitin into its deacetylated—and biotechnologically versatile—form chitosan. Instead, the abundance of endo-chitinase and chitin-binding protein-encoding genes in healthy octocorals leveled up with those from the surrounding environment but was found to be depleted in necrotic octocoral tissue. Using cultivation-independent, taxonomic assignments of endo-chitinase encoding genes, we unveiled previously unsuspected richness and divergent structures of chitinolytic communities across host-associated and free-living biotopes, revealing putative roles for uncultivatedGammaproteobacteriaandChloroflexisymbionts in chitin processing within sessile marine invertebrates.ConclusionsOur findings suggest that differential chitin degradation pathways, utilization, and turnover dictate the processing of chitin across marine micro-niches and support the hypothesis that inter-species cross-feeding could facilitate the co-existence of chitin utilizers within marine invertebrate microbiomes. We further identified chitin metabolism functions which may serve as indicators of microbiome integrity/dysbiosis in corals and reveal putative novel chitinolytic enzymes in the genusAquimarinathat may find applications in the blue biotechnology sector.

Chitin Degradation Machinery and Secondary Metabolite Profiles in the Marine Bacterium Pseudoalteromonas rubra S4059

Genome mining of pigmented Pseudoalteromonas has revealed a large potential for the production of bioactive compounds and hydrolytic enzymes. The purpose of the present study was to explore this bioactivity potential in a potent antibiotic and enzyme producer, Pseudoalteromonas rubra strain S4059. Proteomic analyses (data are available via ProteomeXchange with identifier PXD023249) indicated that a highly efficient chitin degradation machinery was present in the red-pigmented P. rubra S4059 when grown on chitin. Four GH18 chitinases and two GH20 hexosaminidases were significantly upregulated under these conditions. GH19 chitinases, which are not common in bacteria, are consistently found in pigmented Pseudoalteromonas, and in S4059, GH19 was only detected when the bacterium was grown on chitin. To explore the possible role of GH19 in pigmented Pseudoalteromonas, we developed a protocol for genetic manipulation of S4059 and deleted the GH19 chitinase, and compared phenotypes of the mutant and wild type. However, none of the chitin degrading ability, secondary metabolite profile, or biofilm-forming capacity was affected by GH19 deletion. In conclusion, we developed a genetic manipulation protocol that can be used to unravel the bioactive potential of pigmented pseudoalteromonads. An efficient chitinolytic enzyme cocktail was identified in S4059, suggesting that this strain could be a candidate with industrial potential.

Physiological Characterization of the Chitin Synthase A Gene Responsible for Biosynthesis of Cuticle Chitin in Culex Pipiens Pallens (Diptera: Culicidae)

Background: The pathogens transmitted by mosquitoes (Culex pipiens pallens) to humans and animals cause several emerging and resurgent infectious diseases. Increasing insecticide resistance requires rational action to control the target vector population. Chitin is indispensable for insect growth and development and absent from vertebrates and higher plants. Chitin synthase A (CHSA) represents a crucial enzyme in chitin synthesis; therefore, identifying and characterizing how CHSA determines the chitin content might help with novel vector control strategies. Results: The injection of small interfering RNA targeting CHSA (siCHSA) to knock down CHSA transcripts of in larval, pupal, and adult stages, showed different lethal phenotypes. In the larval and pupal stages, CHSA knockdown prevented larval molting, pupation, and adult eclosion, and affected the production of chitin and chitin degradation, which resulted in an ecdysis defect phenotype of mosquitoes. In the adult stage, it also affected the laminar organization of mesoderm and the formation of pseudo orthogonally large fibers of the endoderm. Conclusion: The present study provides a systematic and comprehensive description of the effects of CHSA on morphogenesis and metamorphosis. The results showed that CHSA not only affects chitin synthesis during molting, but also might be involved in chitin degradation. Our result further showed that CHSA is important for the structural integrity of the adult mosquito cuticle.