Advances in extraction, utilization, and development of chitin/chitosan and its derivatives from shrimp shell waste.

Shrimp consumption is in great demand among the seafood used globally. However, this expansion has resulted in the substantial generation and disposal of shrimp shell waste. Through literature search, it has been observed that since 2020, global scholars have shown unprecedented interest in shrimp shell waste and its chitin/chitosan. However, these new insights lack corresponding and comprehensive summarization and analysis. Therefore, this article provides a detailed review of the extraction methods, applications, and the latest research developments on chitin/chitosan from shrimp shells, including micro-nano derivatives, from 2020 to the present. The results indicate that chemical extraction remains the primary technique for the extraction and preparation of chitin/chitosan from shrimp shells. With further refinement and development, adjusting parameters in the chemical extraction process or employing auxiliary techniques such as microwave and radiation enable the customization of target products with different characteristics (e.g., deacetylation degree, molecular weight, and degree of acetylation) according to specific needs. Additionally, in pursuit of environmentally friendly, efficient, and gentle extraction processes, recent research has shifted toward microbial fermentation and green solvent methods for chitin/chitosan extraction. Beyond the traditional antibacterial, film-forming, and encapsulation functionalities, research into the applications of chitosan in biomedical, food processing, new materials, water treatment, and adsorption fields is gradually deepening. Chitin/chitosan derivatives and their modified products have also been a focal point of research in recent years. However, with the rapid expansion, the future development of chitin/chitosan and its derivatives still faces challenges related to the unclear mechanism of action and the complexities associated with industrial scale-up.

Chitin and Its Derivative Chitosan: Distribution in Nature, Applications, and Technology Research (A Review).

The present review highlights the chitin/chitosan as biopolymers that are promising for biomedical research development. Our goal was to assess the potential for practical use of these biopolymers and to summarize information on traditional and innovative technologies for their production and purification. The widespread occurrence of chitin and chitosan in nature as well as the unique chemical and biological properties of chitosan are reasons of growing interest in the use of the latter in several pharmaceutical fields. The main stages of chitin extraction and its further modification into chitosan are deproteinization, demineralization, deacetylation, and the main methods of chitosan purification are filtration, dialysis and reprecipitation. The profitability of the production of chitin/chitosan from crustaceans and edible mushrooms is approximately at the same level. The cost of mushroom products can be reduced by using agricultural or forestry waste as nutrient substrates. This makes the use of fungi as sources of chitin/chitosan in forested regions a rather promising issue.

Exploring the purity of chitin from crustacean sources using deep eutectic solvents: A machine learning approach.

OBJECTIVE: Chitin a natural polymer is abundant in several sources such as shells of crustaceans, mollusks, insects, and fungi. Several possible attempts have been made to recover chitin because of its importance in biomedical applications in various forms such as hydrogel, nanoparticles, nanosheets, nanowires, etc. Among them, deep eutectic solvents have gained much consideration because of their eco-friendly and recyclable nature. However, several factors need to be addressed to obtain a pure form of chitin with a high yield. The development of an innovative system for the production of quality chitin is of prime importance and is still challenging. METHODS: The present study intended to develop a novel and robust approach to investigate chitin purity from various crustacean shell wastes using deep eutectic solvents. This investigation will assist in envisaging the important influencing parameters to obtain a pure form of chitin via a machine learning approach. Different machine learning algorithms have been proposed to model chitin purity by considering the enormous experimental dataset retrieved from previously conducted experiments. Several input variables have been selected to assess chitin purity as the output variable. RESULTS: The statistical criteria of the proposed model have been critically investigated and it was observed that the results indicate XGBoost has the maximum predictive accuracy of 0.95 compared with other selected models. The RMSE and MAE values were also minimal in the XGBoost model. In addition, it revealed better input variables to obtain pure chitin with minimal processing time. CONCLUSION: This study validates that machine learning paves the way for complex problems with substantial datasets and can be an inexpensive and time-saving model for analyzing chitin purity from crustacean shells.

Quantification of Allergic Crustacean Tropomyosin Using Shared Signature Peptides in Processed Foods with a Mass Spectrometry-Based Proteomic Strategy.

Crustacean shellfish are major allergens in East Asia. In the present study, a major allergic protein in crustaceans, tropomyosin, was detected accurately using multiple reaction monitoring mode-based mass spectrometry, with shared signature peptides identified through proteomic analysis. The peptides were deliberately screened through thermal stability and enzymatic digestion efficiency to improve the suitability and accuracy of the developed method. Finally, the proposed method demonstrated a linear range of 0.15 to 30 mgTM/kgfood (R2 > 0.99), with a limit of detection of 0.15 mgTM/kg food and a limit of quantification of 0.5mgTM/kgfood and successfully applied to commercially processed foods, such as potato chips, biscuits, surimi, and hot pot seasonings, which evidenced the applicability of proteomics-based methodology for food allergen analysis.

Nanochitosan from crustacean and mollusk byproduct: Extraction, characterization, and applications in the food industry.

Crustaceans and mollusks are widely consumed around the world due to their delicacy and nutritious value. During the processing, only 30-40 % of these shellfish are considered edible, while 70-60 % of portions are thrown away as waste or byproduct. These byproducts harbor valuable constituents, notably chitin. This chitin can be extracted from shellfish byproducts through chemical, microbial, enzymatic, and green technologies. However, chitin is insoluble in water and most of the organic solvents, hampering its wide application. Hence, chitin is de-acetylated into chitosan, which possesses various functional applications. Recently, nanotechnology has proven to improve the surface area and numerous functional properties of metals and molecules. Further, the nanotechnology principle can be extended to nanochitosan formation. Therefore, this review article centers on crustaceans and mollusks byproduct utilization for chitosan, its nano-formation, and their food industry applications. The extensive discussion has been focused on nanochitosan formation, characterization, and active site modification. Lastly, nanochitosan applications in various food industries, including biodegradable food packaging, fat replacer, bioactive compound carrier, and antimicrobial agent have been reported.

Surface defects due to bacterial residue on shrimp shell.

The changes in the surface chemistry and morphological structure of chitin forms obtained from shrimp shells (ShpS) with and without microorganisms were evaluated. Total mesophilic aerobic bacteria (TMAB), estimated Pseudomonas spp. and Enterococcus spp. were counted in Shp-S by classical cultural counting on agar medium, where the counts were 6.56 ± 0.09, 6.30 ± 0.12, and 3.15 ± 0.03 CFU/g, respectively. Fourier Transform Infrared (FTIR) Spectroscopy and Scanning Electron Microscopy (SEM)/Energy dispersed X-ray (EDX) were used to assess the surface chemistry/functional groups and morphological structure for ChTfree (non-microorganism), and ChTmo (with microorganisms). ChTfree FTIR spectra presented a detailed chitin structure by OH, NH, and CO stretching vibrations, whereas specific peaks of chitin could not be detected in ChTmo. Major differences were also found in SEM analysis for ChTfree and ChTmo. ChTfree had a flat, prominent micropore, partially homogeneous structure, while ChTmo had a layered, heterogeneous, complex dense fibrous, and lost pores form. The degree of deacetylation was calculated for ChTfree and ChTmo according to FTIR and EDX data. The results suggest that the degree of deacetylation decreases in the presence of microorganisms, affecting the production of beneficial components negatively. The findings were also supported by the molecular docking model.

A crustacean neuropeptide spectral library for data-independent acquisition (DIA) mass spectrometry applications.

Neuropeptides have tremendous potential for application in modern medicine, including utility as biomarkers and therapeutics. To overcome the inherent challenges associated with neuropeptide identification and characterization, data-independent acquisition (DIA) is a fitting mass spectrometry (MS) method of choice to achieve sensitive and accurate analysis. It is advantageous for preliminary neuropeptidomic studies to occur in less complex organisms, with crustacean models serving as a popular choice due to their relatively simple nervous system. With spectral libraries serving as a means to interpret DIA-MS output spectra, and Cancer borealis as a model of choice for neuropeptide analysis, we performed the first spectral library mapping of crustacean neuropeptides. Leveraging pre-existing data-dependent acquisition (DDA) spectra, a spectral library was built using PEAKS Online. The library is comprised of 333 unique neuropeptides. The identification results obtained through the use of this spectral library were compared with those achieved through library-free analysis of crustacean brain, pericardial organs (PO), and thoracic ganglia (TG) tissues. A statistically significant increase (Student's t-test, P value < 0.05) in the number of identifications achieved from the TG data was observed in the spectral library results. Furthermore, in each of the tissues, a distinctly different set of identifications was found in the library search compared to the library-free search. This work highlights the necessity for the use of spectral libraries in neuropeptide analysis, illustrating the advantage of spectral libraries for interpreting DIA spectra in a reproducible manner with greater neuropeptidomic depth.

Production of chitosan from shrimp shell using ultrasound followed by subcritical water hydrolysis.

The interest in shrimp shell valorization has been growing in line with sustainability goals. Therefore, the main objective of this study was to obtain chitosan from shrimp shell using ultrasound followed by subcritical water treatment. Ultrasonication of shells was performed at 600 and 1200 W for 5 min. Then, shells were hydrolyzed at 140-260 °C and 50 bar for 10-60 min followed by demineralization using citric acid, bleaching using hydrogen peroxide and deacetylation using sodium hydroxide solution. The highest deproteination (80.93 %) was obtained by ultrasonication at 1200 W/5 min followed by subcritical water hydrolysis at 260 °C/50 bar/60 min, where the residue with a yield of 10.56 %, whiteness index of 60.42, degree of deacetylation of 64.27 %, relative crystallinity of 32.66 % and similar functional groups to the commercial sample was obtained. These results indicated that the combination of ultrasound with subcritical water is promising to valorize shrimp shell towards production of value-added compounds.

The Preparation and Physiochemical Characterization of Tenebrio molitor Chitin Using Alcalase.

Chitin is mostly produced from crustaceans, but it is difficult to supply raw materials due to marine pollution, and the commonly used chemical chitin extraction method is not environmentally friendly. Therefore, this study aims to establish a chitin extraction process using enzymes and to develop edible insect-derived chitin as an eco-friendly new material. The response surface methodology (RSM) was used to determine the optimal conditions for enzymatic hydrolysis. The optimal conditions for enzymatic hydrolysis by RSM were determined to be the substrate concentration (7.5%), enzyme concentration (80 µL/g), and reaction time (24 h). The solubility and DDA of the mealworm chitosan were 45% and 37%, respectively, and those of the commercial chitosan were 61% and 57%, respectively. In regard to the thermodynamic properties, the exothermic peak of mealworm chitin was similar to that of commercial chitin. In the FT-IR spectrum, a band was observed in mealworm chitin corresponding to the C=O of the NHCOCH3 group at 1645 cm-1, but this band showed low-intensity C=O in the mealworm chitosan due to deacetylation. Collectively, mealworm chitosan shows almost similar physical and chemical properties to commercial chitosan. Therefore, it is shown that an eco-friendly process can be introduced into chitosan production by using enzyme-extracted mealworms for chitin/chitosan production.

Chitosan Based Materials in Cosmetic Applications: A Review.

This review provides a report on the properties and recent advances in the application of chitosan and chitosan-based materials in cosmetics. Chitosan is a polysaccharide that can be obtained from chitin via the deacetylation process. Chitin most commonly is extracted from cell walls in fungi and the exoskeletons of arthropods, such as crustaceans and insects. Chitosan has attracted significant academic interest, as well as the attention of the cosmetic industry, due to its interesting properties, which include being a natural humectant and moisturizer for the skin and a rheology modifier. This review paper covers the structure of chitosan, the sources of chitosan used in the cosmetic industry, and the role played by this polysaccharide in cosmetics. Future aspects regarding applications of chitosan-based materials in cosmetics are also mentioned.

Simultaneous determination of five metabolites of nitrofurans including the metabolite nifursol in shrimp and fish by UPLC- MS/MS: in-house method validation according to commission implementing regulation (EU) 2021/808.

For the simultaneous identification and quantification of five nitrofurans metabolites in farmed shrimp and fish, 3-amino-2-oxazolidinone (AOZ), 3-amino-5-morpholinomethyl-2-oxazolidinone (AMOZ), 1-aminohydantoine (AHD), semicarbazide (SEM), and 3,5-dinitrosalicylic acid hydrazide (DNSH), an accurate, precise, and specific method was developed. The mixture of water and methanol (60/40; v/v) was found to be the final optimised solvent for injection. The analytical run duration was 7 min, and the mobile phase included 2 mM methanol and ammonium formate. The new reference point for action (RPA) of 0.50 µg kg-1 as per EC/1871/2019 was taken into consideration and evaluated for the performance characteristics as per the CIR (EC)/2021/808 criteria. Specificity, relative retention time (≤0.25%) relative ion ratio (≤40%), linearity (0.25 to 2.0 µg kg-1), trueness (between 82.8 and 118.1%), repeatability (RSDr ≤14%), within lab reproducibility (RSDwr ≤16.9%), CCα (0.32-0.36 µg kg-1), ruggedness and relative matrix effect (≤14.26%) achieved acceptable values.

Exploring Saduria entomon (Crustacea Isopoda) as a New Source for Chitin and Chitosan Isolation.

Chitin and chitosan demand is growing very fast due to interest from industries such as pharmaceutical, cosmetic, agricultural and others. New sources for chitin and chitosan isolation are being extensively searched to fulfil this demand. In this paper, Saduria entomon a Baltic benthic crustacean, is evaluated as a source for chitin and chitosan isolation. Chitin and chitosan yield from S. entomon were 14.8 and 8.2%, respectively, in a similar range to other sources. Samples were characterized in terms of physicochemical properties (acetylation degree, molecular weight, thermal stability, and crystallinity) and two biological properties, antimicrobial activity and antioxidant activity were evaluated. Chitosan S. entomon exhibited antimicrobial activity against S. aureus but not against E. coli. An antioxidant activity of 20.98 TROLOX µmol equivalent/g polymer was detected for the chitosan sample. These properties are very promising for the use of this organism as a source for chitin and chitosan isolation in the biomedical field.

A Review of the Chemical Extraction of Chitosan from Shrimp Wastes and Prediction of Factors Affecting Chitosan Yield by Using an Artificial Neural Network.

There are two viable options to produce shrimp shells as by-product waste, either within the shrimp production phases or when the shrimp are peeled before cooking by the end user. This waste is considered a double-edged sword, as it is possible to be either a source of environmental pollution, through dumping and burning, or a promising source from which to produce chitosan as a biodegradable, biocompatible biopolymer which has a variety of agricultural, industrial, and biomedical applications. Chitosan is a deacetylated form of chitin that can be chemically recovered from shrimp shells through the three sequential stages of demineralization, deproteinization, and deacetylation. The main aim of this review paper is to summarize the recent literature on the chemical extraction of chitosan from shrimp shells and to represent the physicochemical properties of chitosan extracted from shrimp shells in different articles, such as chitosan yield, moisture content, solubility, ash content, and degree of deacetylation. Another aim is to analyze the influence of the main predictors of the chemical extraction stages (demineralization, deproteinization, and deacetylation) on the chitosan yield percentage by using a multilayer perceptron artificial neural network. This study showed that the deacetylation alkali concentration is the most crucial parameter, followed by the concentrations of acid and alkali of demineralization and deproteinization, respectively. The current review was conducted to be used in prospective studies for optimizing the chemical extraction of chitosan from shrimp wastes.

The impact of the different types of acid solution on the extraction and adsorption performance of chitin from shrimp shell waste.

The properties of chitin-based adsorbents varied among studies since they are influenced by different factors, such as the types of base and acid used to extract the chitin. Therefore, this works aimed to investigate the impact of four different acid solutions on the extraction and properties of chitin from shrimp shell waste, and to evaluate the adsorption performance of the obtained chitin on removing dye from an aqueous solution. The result showed that H2SO4, HCl, and HNO3 could remove high minerals from the shrimp shell, while the effect of CH3COOH was inferior. The Fourier Transform Infrared (FTIR) and X-ray diffraction (XRD) indicated that the extracted chitin was α-amorphous structure, regardless of the type of acid solution. However, the type of acid solution influenced the crystallinity index of the extracted chitin. The Scanning Electron Microscope (SEM) showed both fibrillar material and porous structures. In addition, the chitin extracted through demineralization using H2SO4 was more effective in removing RBBR dye from aqueous solution, followed by HCl, HNO3, and the last, CH3COOH treatment. The performances of chitin-based adsorbent could be attributed to the strength of acid solution used to remove mineral during the extraction process and the obtained pore structures.

Exploring new marine bacterial species, Alcaligenes faecalis Alca F2018 valued for bioconversion of shrimp chitin to chitosan for concomitant biotechnological applications.

The exploitation of chitinous materials seems to be an infinite treasure. To this end, using shellfish waste as the sole carbon/nitrogen source solves environmental challenges while lowering microbial chitinase production costs. Bioconversion of shellfish chitin wastes such as shrimp shells has recently been investigated for the production of enzymes and bioactive materials in order to maximize the utilization of chitin-containing seafood processing wastes. In this study, the bioconversion of chitin to chitosan by Alcaligenes faecalis Alca F2018 revealed the highest chitin deacetylase (CDA) activity of 40.6 U/µg. The resulted low Km and high Vmax values explain the high affinity of the purified CDA to the p-nitroacetanilide substrate. CDA with a molecular weight of 66 KDa was purified from F2018 strain, with a 14.5% yield. FT-IR revealed distinct chitosan peaks and XRD revealed that chitosan samples had lower crystallinity than chitin. TGA analysis revealed that the recovered chitosan samples were more thermally stable. The deacetylation degree percentages of the produced chitosan are in the same range as that of the commercial chitosan, suggesting the promising potential of A. faecalis Alca F2018 to utilize shrimp shells in their raw form in the fermentation media based on its CDA enzyme activity.

Marine paralytic shellfish toxins: chemical properties, mode of action, newer analogues, and structure-toxicity relationship.

Up to the end of 2020Every year, the appearance of marine biotoxins causes enormous socio-economic damage worldwide. Among the major groups of biotoxins, paralytic shellfish toxins, comprising saxitoxin and its analogues (STXs), are the ones that cause the most severe effects on humans, including death. However, the knowledge that currently exists on their chemistry, properties and mode of toxicological action is disperse and partially outdated. This review intends to systematically compile the dispersed information, updating and complementing it. With this purpose, it addresses several aspects related to the molecular structure of these toxins. Special focus is given to the bioconversion reactions that may occur in the different organisms (dinoflagellates, bivalves, and humans) and the possible mediators involved. A critical review of the most recently discovered analogues, the M-series toxins, is presented. Finally, a deep discussion about the relationship between the molecular structure (e.g., effect of the substituting groups and the net charge of the molecules) and the toxic activity of these molecules is performed, proposing the concept of "toxicological traffic light" based on the toxicity equivalency factors (TEFs).

A Comparative Analysis of Conventional and Deep Eutectic Solvent (DES)-Mediated Strategies for the Extraction of Chitin from Marine Crustacean Shells.

Chitin, the second most abundant biopolymer on earth, is utilised in a wide range of applications including wastewater treatment, drug delivery, wound healing, tissue engineering, and stem cell technology among others. This review compares the most prevalent strategies for the extraction of chitin from crustacean sources including chemical methods that involve the use of harsh solvents and emerging methods using deep eutectic solvents (DES). In recent years, a significant amount of research has been carried out to identify and develop environmentally friendly processes which might facilitate the replacement of problematic chemicals utilised in conventional chemical extraction strategies with DES. This article provides an overview of different experimental parameters used in the DES-mediated extraction of chitin while also comparing the purity and yields of associated extracts with conventional methods. As part of this review, we compare the relative proportions of chitin and extraneous materials in different marine crustaceans. We show the importance of the species of crustacean shell in relation to chitin purity and discuss the significance of varying process parameters associated with different extraction strategies. The review also describes some recent applications associated with chitin. Following on from this review, we suggest recommendations for further investigation into chitin extraction, especially for experimental research pertaining to the enhancement of the "environmentally friendly" nature of the process. It is hoped that this article will provide researchers with a platform to better understand the benefits and limitations of DES-mediated extractions thereby further promoting knowledge in this area.

A g-Type Lysozyme from Deep-Sea Hydrothermal Vent Shrimp Kills Selectively Gram-Negative Bacteria.

Lysozyme is a key effector molecule of the innate immune system in both vertebrate and invertebrate. It is classified into six types, one of which is the goose-type (g-type). To date, no study on g-type lysozyme in crustacean has been documented. Here, we report the identification and characterization of a g-type lysozyme (named LysG1) from the shrimp inhabiting a deep-sea hydrothermal vent in Manus Basin. LysG1 possesses conserved structural features of g-type lysozymes. The recombinant LysG1 (rLysG1) exhibited no muramidase activity and killed selectively Gram-negative bacteria in a manner that depended on temperature, pH, and metal ions. rLysG1 bound target bacteria via interaction with bacterial cell wall components, notably lipopolysaccharide (LPS), and induced cellular membrane permeabilization, which eventually caused cell lysis. The endotoxin-binding capacity enabled rLysG1 to alleviate the inflammatory response induced by LPS. Mutation analysis showed that the bacterial binding and killing activities of rLysG1 required the integrity of the conserved α3 and 4 helixes of the protein. Together, these results provide the first insight into the activity and working mechanism of g-type lysozyme in crustacean and deep-sea organisms.

Cytochalasin Derivatives from the Endozoic Curvularia verruculosa CS-129, a Fungus Isolated from the Deep-Sea Squat Lobster Shinkaia crosnieri Living in the Cold Seep Environment.

A new cytochalasin dimer, verruculoid A (1), three new cytochalasin derivatives, including 12-nor-cytochalasin F (2), 22-methoxycytochalasin B6 (3), and 19-hydroxycytochalasin B (4), and 20-deoxycytochalasin B (5), a synthetic product obtained as a natural product for the first time, together with four known analogues (6-9), were isolated and identified from the culture extract of Curvularia verruculosa CS-129, an endozoic fungus obtained from the inner fresh tissue of the deep-sea squat lobster Shinkaia crosnieri, which was collected from the cold seep area of the South China Sea. Structurally, verruculoid A (1) represents the first cytochalasin homodimer containing a thioether bridge, while 12-nor-cytochalasin F (2) is the first 12-nor-cytochalasin derivative. Their structures were elucidated by detailed interpretation of the NMR spectroscopic and mass spectrometric data. X-ray crystallographic analysis and ECD calculations confirmed their structures and absolute configurations. Compound 1 displayed activity against the human pathogenic bacterium Escherichia coli (MIC = 2 µg/mL), while compounds 4, 8, and 9 showed cytotoxicity against three tumor cell lines (HCT-116, HepG-2, and MCF-7) with IC50 values from 5.2 to 12 µM. The structure-activity relationship was briefly discussed.

Structure and mineralization of the spearing mantis shrimp (Stomatopoda; Lysiosquillina maculata) body and spike cuticles.

Stomatopoda is a crustacean order including sophisticated predators called spearing and smashing mantis shrimps that are separated from the well-studied Eumalacotraca since the Devonian. The spearing mantis shrimp has developed a spiky dactyl capable of impaling fishes or crustaceans in a fraction of second. In this high velocity hunting technique, the spikes undergo an intense mechanical constraint to which their exoskeleton (or cuticle) has to be adapted. To better understand the spike cuticle internal architecture and composition, electron microscopy, X-ray microanalysis and Raman spectroscopy were used on the spikes of 7 individuals (collected in French Polynesia and Indonesia), but also on parts of the body cuticle that have less mechanical stress to bear. In the body cuticle, several specificities linked to the group were found, allowing to determine the basic structure from which the spike cuticle has evolved. Results also highlighted that the body cuticle of mantis shrimps could be a model close to the ancestral arthropod cuticle by the aspect of its biological layers (epi- and procuticle including exo- and endocuticle) as well as by the Ca-carbonate/phosphate mineral content of these layers. In contrast, the spike cuticle exhibits a deeply modified organization in four functional regions overprinted on the biological layers. Each of them has specific fibre arrangement or mineral content (fluorapatite, ACP or phosphate-rich Ca-carbonate) and is thought to assume specific mechanical roles, conferring appropriate properties on the entire spike. These results agree with an evolution of smashing mantis shrimps from primitive stabbing/spearing shrimps, and thus also allowed a better understanding of the structural modifications described in previous studies on the dactyl club of smashing mantis shrimps.