Proteases Production and Chitin Preparation from the Liquid Fermentation of Chitinous Fishery By-Products by Paenibacillus elgii

Chitinous fishery by-products have great application in the production of various bioactive compounds. In this study, Paenibacillus elgii TKU051, a protease-producing bacterial strain, was isolated using a medium containing 1% squid pens powder (SPP) as the sole carbon/nitrogen (C/N) source. P. elgii TKU051 was found to produce at least four proteases with molecular weights of 100 kDa, 57 kDa, 43 kDa, and 34 kDa (determined by the gelatin zymography method). A P. elgii TkU051 crude enzyme cocktail was optimally active at pH 6–7 and 60 °C. The 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity and α-glucosidase inhibitory activity of the hydrolysates obtained from the hydrolysis of shrimp shell powder, shrimp head powder, shrimp meat powder, fish head powder and soya bean powder catalyzed by the P. elgii TkU051 crude enzyme cocktail were also evaluated. P. elgii TKU051 exhibited a high deproteinization capacity (over 94%) on different kinds of shrimp waste (shrimp heads and shells; fresh and cooked shrimp waste; shrimp waste dried by oven and lyophilizer), and the Fourier-transform infrared spectroscopy profile of the chitin obtained from the deproteinization process displayed the characteristic of chitin. Finally, the obtained chitin exhibited an effect comparable to commercial chitin in terms of adsorption against Congo Red (90.48% and 90.91%, respectively). Thus, P. elgii TKU051 showed potential in the reclamation of chitinous fishery by-products for proteases production and chitin extraction.     

The Effect of Molecular Weight on the Antimicrobial Activity of Chitosan from Loligo opalescens for Food Packaging Applications

The growing requirement for sustainable processes has boosted the development of biodegradable plastic-based materials incorporating bioactive compounds obtained from waste, adding value to these products. Chitosan (Ch) is a biopolymer that can be obtained by deacetylation of chitin (found abundantly in waste from the fishery industry) and has valuable properties such as biocompatibility, biodegradability, antimicrobial activity, and easy film-forming ability. This study aimed to produce and characterize poly(lactic acid) (PLA) surfaces coated with β-chitosan and β-chitooligosaccharides from a Loligo opalescens pen with different molecular weights for application in the food industry. The PLA films with native and depolymerized Ch were functionalized through plasma oxygen treatment followed by dip-coating, and their physicochemical properties were assessed by Fourier-transform infrared spectroscopy, X-ray diffraction, water contact angle, and scanning electron microscopy. Their antimicrobial properties were assessed against Escherichia coli and Pseudomonas putida, where Ch-based surfaces reduced the number of biofilm viable, viable but nonculturable, and culturable cells by up to 73%, 74%, and 87%, respectively, compared to PLA. Biofilm growth inhibition was confirmed by confocal laser scanning microscopy. Results suggest that Ch films of higher molecular weight had higher antibiofilm activity under the food storage conditions mimicked in this work, contributing simultaneously to the reuse of marine waste.     

Biodegradation and Prospect of Polysaccharide from Crustaceans

Marine crustacean waste has not been fully utilized and is a rich source of chitin. Enzymatic degradation has attracted the wide attention of researchers due to its unique biocatalytic ability to protect the environment. Chitosan (CTS) and its derivative chitosan oligosaccharides (COSs) with various biological activities can be obtained by the enzymatic degradation of chitin. Many studies have shown that chitosan and its derivatives, chitosan oligosaccharides (COSs), have beneficial properties, including lipid-lowering, anti-inflammatory and antitumor activities, and have important application value in the medical treatment field, the food industry and agriculture. In this review, we describe the classification, biochemical characteristics and catalytic mechanisms of the major degrading enzymes: chitinases, chitin deacetylases (CDAs) and chitosanases. We also introduced the technology for enzymatic design and modification and proposed the current problems and development trends of enzymatic degradation of chitin polysaccharides. The discussion on the characteristics and catalytic mechanism of chitosan-degrading enzymes will help to develop new types of hydrolases by various biotechnology methods and promote their application in chitosan.     

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.     

贵州山区生态茶园不同凋落物木质素、纤维素分解特征

采用凋落物分解袋法,以贵州久安生态茶园内云南樟（Cinnamomum glanduliferum）、光皮桦（Betula luminifera）、杉木（Cunninghamia lanceolata）和马尾松（Pinus massoniana）叶凋落物以及茶树（Camellia sinensis）修剪物为研究对象,分析5种单一凋落物类型,以及9种混合凋落物类型（茶树-云南樟、茶树-马尾松、茶树-杉木、茶树-光皮桦、杉木-光皮桦、云南樟-光皮桦、云南樟-马尾松、云南樟-杉木、光皮桦-马尾松）分解速率和木质素、纤维素释放特征及差异性。结果表明,单一凋落物类型中,阔叶树种凋落物分解速率大于针叶树种;阔叶树种凋落物木质素含量总体上呈现增加趋势,而针叶树种总体上则呈现下降趋势;凋落物纤维素含量总体上呈现下降趋势。单一凋落物纤维素释放规律与混合凋落物纤维素释放一致,均为直接释放;单一凋落物木质素释放存在3种规律;混合凋落物木质素释放规律有5种。分析9种混合凋落物分解特征,44.4%的混合凋落物混合分解表现为加和效应,11.2%的混合分解表现为拮抗效应,44.4%的混合分解表现为协同效应;22.2%的混合凋落物木质素的释放表现为促进释放,55.6%的混合凋落物木质素释放表现为促进富集;22.2%的混合凋落物纤维素释放表现为促进释放;11.1%的混合凋落物纤维素释放表现为促进富集。研究结果可为维持茶园肥力提供理论依据。     

Chitin and Chitosan as Multipurpose Natural Polymers for Groundwater Arsenic Removal and As2O3 Delivery in Tumor Therapy

Chitin and chitosan are natural polysaccharide polymers. These polymers have been used in several agricultural, food protection and nutraceutical applications. Moreover, chitin and chitosan have been also used in biomedical and biotechnological applications as drug delivery systems or in pharmaceutical formulations. So far, there are only few studies dealing with arsenic (As) removal from groundwater using chitin or chitosan and no evidence of the use of these natural polymers for arsenic trioxide (As₂O₃) delivery in tumor therapy. Here we suggest that chitin and/or chitosan might have the right properties to be employed as efficient polymers for such applications. Besides, nanotechnology offers suitable tools for the fabrication of novel nanostructured materials of natural origin. Since different nanostructured materials have already been employed successfully in various multidisciplinary fields, we expect that the integration of nanotechnology and natural polymer chemistry will further lead to innovative applications for environment and medicine.     

Bioproduction of Chitooligosaccharides: Present and Perspectives

Chitin and chitosan oligosaccharides (COS) have been traditionally obtained by chemical digestion with strong acids. In light of the difficulties associated with these traditional production processes, environmentally compatible and reproducible production alternatives are desirable. Unlike chemical digestion, biodegradation of chitin and chitosan by enzymes or microorganisms does not require the use of toxic chemicals or excessive amounts of wastewater. Enzyme preparations with chitinase, chitosanase, and lysozymeare primarily used to hydrolyze chitin and chitosan. Commercial preparations of cellulase, protease, lipase, and pepsin provide another opportunity for oligosaccharide production. In addition to their hydrolytic activities, the transglycosylation activity of chitinolytic enzymes might be exploited for the synthesis of desired chitin oligomers and their derivatives. Chitin deacetylase is also potentially useful for the preparation of oligosaccharides. Recently, direct production of oligosaccharides from chitin and crab shells by a combination of mechanochemical grinding and enzymatic hydrolysis has been reported. Together with these, other emerging technologies such as direct degradation of chitin from crustacean shells and microbial cell walls, enzymatic synthesis of COS from small building blocks, and protein engineering technology for chitin-related enzymes have been discussed as the most significant challenge for industrial application.     

不同肥力土壤中分解几丁质微生物代谢产物对玉米的促生效果研究

从吉林省不同玉米栽培区土壤中以胶状几丁质为碳源分离筛选分解几丁质微生物,最终筛选出功能菌株并命名为CC-1,采用16 Sr RNA序列扩增法鉴定为芽孢杆菌Bacillus sp,通过SDS-PAGE分析法进一步确定该菌株的几丁质酶生产能力和几丁质酶的蛋白分子量为30 k Da。通过盆栽试验进行微生物肥料对不同肥力中的玉米促生效果的研究,结果表明,以CC-1菌株生产的微生物肥料效果受土壤的基础肥力状况影响较大,土壤的肥力状况与微生物的增产效应成反比,相对贫瘠的土壤对微生物处理的响应较为敏感,对玉米的生物学性状及产量表现出了一定的促进作用,增产达到了13.08%。     

Carboxymethyl lignin as stabilizing agent in aqueous ceramic suspensions

Identifying new uses for residues of industries that process large quantities of biomass, as in bioethanol production, is essential for a sustainable development with reduced impact on the environment, which is the reason why many efforts have been devoted to find noble uses for lignins. In this study, a lignin obtained from sugarcane bagasse in a bioethanol producing plant was carboxymethylated to yield the water-soluble carboxymethyl lignin (CML), which was then used as stabilizing agent in aqueous alumina (Al₂O₃) suspensions. CML had a degree of substitution 0.46 ± 0.01, in relation to the C9 unit of lignin, and behaved as a polyelectrolyte in a large pH range owing to the dissociation of carboxylic groups. The action of CML as stabilizing agent of alumina aqueous suspensions was investigated using viscometry, zeta potential, and photon correlation spectroscopy (PCS) measurements, mainly as a function of pH and time. Overall, the results showed that CML had a good performance as a deflocculating agent, because it led to dispersions with low viscosity and small change in particle size as a function of time. The positive effect from the addition of CML was confirmed in the morphological features of the material obtained from the alumina suspensions after elimination of water, as indicated by scanning electron microscopy. The stabilization of alumina suspensions afforded by CML opens the way for similar applications of modified lignins, whose electrical and structural properties may be tuned for specific uses in various industries, including the ceramic industry.     

Applicability of Industrial Sisal Fiber Waste Derived Activated Carbon for the Adsorption of Volatile Organic Compounds (VOCs)

Agricultural waste produced by the industry is a huge threat for the global environment. Utilization of agricultural waste is necessary and there is an urgent need to develop new techniques to solve this important problem. The main objective of this research was to evaluate the applicability of the activated carbon (AC) derived from industrial sisal fiber waste as passive samplers in monitoring toluene by comparing them to industry standard wafer and granular activated carbon (GAC). Carbonization time and ball milling effect on sisal fiber derived AC sample adsorption properties were investigated. Toluene adsorption isotherms were used to predict toluene adsorption capacities. Surface characteristics including surface area and pore volume were used to determine the relationships between them and adsorption capacity. Even though prepared AC samples have mesoporous structure, commercial samples have microporous structure. Surface area from 1245 m²/g to 1297 m²/g and toluene adsorption capacity from 21.4 % to 26.6 % was improved by increasing carbonization time from 1 h to 3 h at 650 °C carbonization temperature and 94.4 ml/min flow rate. Conversely, ball milling technique had negative effect by decreasing the surface area (674 m²/g) and the adsorption capacity of toluene (12.27 %). It is concluded that industrial sisal fiber waste have great potential as a precursor of AC for application in passive monitoring against toluene, particularly the produced mesoporous AC with 3 h carbonization time performs higher adsorption capacity (26.6 %) than commercially available microporous passive sampler (24.1 %) and GAC (22.8 %).     

Co-composting of lignin to build up humic substances—Strategies in waste management to improve compost quality

Co-composting of different lignin sources with biogenic waste from the separate collection in Vienna was carried out in lab-scale experiments to promote humification as lignin is known to be a precursor molecule of humic substances. Humification is a target to be reached by composting due to the favorable properties for soils and plants. Extractable humic acids have been proved to be a suitable parameter to assess humification. Biogenic waste was used as a reference and basic material for co-composting of lignin. Lignin from different origins and composition was added: microbiologically degraded wood and lignin powder from industrial processes. Preliminary results show that addition of 2–5% of lignin powder could improve humic acid formation. Microbiologically degraded wood did not effect higher humic acid contents within the short time of lab-scale composting.Humic acid determination is based on photometrical investigations. To ensure that humic acid contents were not pretended by the presence of lignin, quality control was carried out using FT-IR spectroscopy and thermal analysis. The results obtained showed that lignin had been integrated in the humic acid molecule.     

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.     

Functional Properties and Dietary Fiber Characterization of Mango Processing By-products (Mangifera indica L., cv Ataulfo and Tommy Atkins)

Several reports have focused on utilization of post-harvest residues of crops, while neglecting those residues produced by mango processing. These residues represent a waste of nutrients and a source of environmental contaminants. Such by-products could be valuable sources of dietary fiber (DF), antioxidant compounds, and single carbohydrates. The aim of this study was to evaluate some functional properties (FP), and the content of DF and polyphenols (PP) of the peel and coarse material obtained from residues during the industrial processing of Ataulfo and Tommy Atkins mangoes. The total dietary fiber (TDF) content was about 225 mg/g and 387 mg/g (dry weight) for the coarse material and the peel, respectively, from which soluble dietary fiber represented 23 and 42 %, respectively. The main neutral sugar identified was rhamnose, especially in peels; the klason lignin (KL) content was 92 mg/g, which highlights the Ataulfo peel (Ataulfo-P) and the Tommy Atkins peel (Tommy Atkins-P). The extractable PP content in Ataulfo-P was higher than in Tommy-Atkins-P, and interesting data for non-extractable PP were obtained in the residues. FP as swelling, water holding, oil holding, and glucose absorption in the residues was studied, obtaining better functional properties when compared to cellulose fiber. The results show that mango industrial by-products, mainly from the Ataulfo-P variety, could be used as ingredients in food products because of their functional properties as well as their DF and PP content.     

吉林人参中木质素含量的测定

目的:建立紫外分光光度法测定人参中木质素含量的方法并研究吉林人参中木质素的含量。方法:采用经典的Klason法和紫外分光光度法。结果:人参木质素特征吸收峰在268nm。紫外分光光度法操作简便、准确、快速,适合测定人参中木质素含量的测定。结论:吉林省不同产地、不同生长年份的人参所含木质素的量是不同的。     

The optimal lignin quantification method to breed for an improved cell wall digestibility in perennial ryegrass

Perennial ryegrass (Lolium perenne) is an important source of protein and energy for dairy cattle. To improve the protein/energy ratio of this forage, focus is now on improving its cell wall digestibility. The in vitro assessment of the digestible fraction of the neutral detergent fibre (NDFD) is a superior method for determining the cell wall digestibility, but requires the use of ruminal fluid, which has a highly variable composition and is often not readily available. As lignin is considered the main cell wall component that impedes NDFD, we investigated whether this “subtrait” could serve as alternative breeding selection criterion to improve NDFD. Therefore, we assessed the accuracy of two lignin quantification methods: van Soest (ADL) and Klason lignin (KL). We also considered KL estimates corrected for the solubilized lignin (total lignin or TL) and non‐solubilized protein (TL'). Although the latter is considered the truest possible lignin content, it was not always the most correlated to NDFD, due to the limited accuracy of protein quantification on lignin residue. TL is most correlated to NDFD and we therefore recommend it for use in conventional breeding if NDFD determination is not a possibility. However, NDFD is still a superior selection criterion, as it combines the effect of several subtraits and not just lignin. For marker–trait association studies, a more accurate estimate of lignin content is more important than a high correlation with NDFD, but also here, TL performs best.     

Protective Effect of Chitin Urocanate Nanofibers against Ultraviolet Radiation

Urocanic acid is a major ultraviolet (UV)-absorbing chromophore. Chitins are highly crystalline structures that are found predominantly in crustacean shells. Alpha-chitin consists of microfibers that contain nanofibrils embedded in a protein matrix. Acid hydrolysis is a common method used to prepare chitin nanofibrils (NFs). We typically obtain NFs by hydrolyzing chitin with acetic acid. However, in the present study, we used urocanic acid to prepare urocanic acid chitin NFs (UNFs) and examined its protective effect against UVB radiation. Hos: HR-1 mice coated with UNFs were UVB irradiated (302 nm, 150 mJ/cm²), and these mice showed markedly lower UVB radiation-induced cutaneous erythema than the control. Additionally, sunburn cells were rarely detected in the epidermis of UNFs-coated mice after UVB irradiation. Although the difference was not as significant as UNFs, the number of sunburn cells in mice treated with acetic acid chitin nanofibrils (ANFs) tended to be lower than in control mice. These results demonstrate that ANFs have a protective effect against UVB and suggest that the anti-inflammatory and antioxidant effects of NFs influence the protective effect of ANFs against UVB radiation. The combination of NFs with other substances that possess UV-protective effects, such as urocanic acid, may provide an enhanced protective effect against UVB radiation.     

Water-resistant Lignin/Poly(vinyl alcohol) Blend Fibers for Removal of Hexavalent Chromium

Lignin is the second most abundant renewable biomass-derived natural resource that has been used to replace traditional petrochemical-based materials. However, fabricating the lignin component into the various forms required for practical application is still challenging. In this work, we fabricated water-resistant lignin/poly(vinyl alcohol) (PVA) blend fibers by wet spinning and glutaraldehyde crosslinking methods. The effect of the lignin/PVA blend ratio and glutaraldehyde crosslinking process on the physicochemical properties of wet-spun lignin/PVA blend fibers were studied using maximum draw ratios, hydrolytic degradation profiles, and mechanical properties. Furthermore, the hexavalent chromium [Cr(VI)] removal behavior of lignin/PVA blend fibers was investigated according to the effect of pH, initial Cr(VI) concentration, and contact time. The wet-spun lignin/PVA blend fiber achieved excellent water stability through glutaraldehyde crosslinking and exhibited notable Cr(VI) adsorption capacity (350.87 mg/g) and good regeneration ability. These findings demonstrate that glutaraldehyde-crosslinked lignin/PVA blend fibers could be promising adsorbents for the remediation of heavy metal species containing textile wastewater.     

Synthesis and characterization of hybrid latexes from soybean oil-based polyurethane and poly(2,2,2-trifluoroethyl methacrylate)

A class of novel fluorine containing core-shell hybrid latexes were obtained from soybean oil-based polyurethanes and poly(2,2,2-trifluoroethyl methacrylate) (PTFEMA) by two-step synthesis under mild reaction conditions. Structural and morphology properties of the resulting hybrid latexs have been characterized by FT-IR and TEM. In addition, thermal properties (DSC, TGA) and coating performance (contact angle, stress-strain curve, and surface free energy) were investigated and discussed. The hybrid latexes exhibit outstanding thermal stability and coating performance. More importantly, through this method, low-valued foods are successfully transformed into high-valued functional materials which bring new solutions for preparing materials from renewable sources.