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.

Chitin Determination in Residual Streams Derived From Insect Production by LC-ECD and LC-MS/MS Methods

Chitin, a biopolymer present in fungi and arthropods, is a compound of interest for various applications, such as in the agricultural and medical fields. With the recently growing interest in the development of insect farming, the availability of chitin-containing residual streams, particularly the molting skins (exuviae), is expected to increase in the near future. For application purposes, accurate quantification of chitin in these insect sources is essential. Previous studies on chitin extraction and quantification often overlooked the purity of the extracted chitin, making the outcomes inconsistent and prone to overestimation. The present study aims to determine chitin content in the exuviae of three insect species mass-reared worldwide: black soldier fly (BSF), mealworm, and house cricket. Chitin was chemically extracted using acid and alkali treatments to remove minerals and proteins. The purity of extracted chitin was evaluated by hydrolyzing the chitin into glucosamine, followed by quantitative determination of the latter using two liquid chromatography methods: electrochemical detection (ECD) and tandem mass spectrometry (MS/MS). Both methods proved accurate and precise, without the need for labor-intensive derivatization steps. Pearson's correlation and Bland-Altman plots showed that the glucosamine determination results obtained by the two methods were comparable, and there is no consistent bias of one approach vs. the other. The chitin content in extracted residues ranged between 7.9 and 18.5%, with the highest amount found in BSF puparium. In summary, the study demonstrated that (1) the residual streams of the insect farming industry have a great potential for utilization as an alternative chitin source, and (2) both LC-ECD and LC-MS/MS are reliable for the quantitative determination of glucosamine in insect chitin.

Antimicrobial Activity of Chemically and Biologically Treated Chitosan Prepared from Black Soldier Fly (Hermetia illucens) Pupal Shell Waste

Globally, the broad-spectrum antimicrobial activity of chitin and chitosan has been widely documented. However, very little research attention has focused on chitin and chitosan extracted from black soldier fly pupal exuviae, which are abundantly present as byproducts from insect-farming enterprises. This study presents the first comparative analysis of chemical and biological extraction of chitin and chitosan from BSF pupal exuviae. The antibacterial activity of chitosan was also evaluated. For chemical extraction, demineralization and deproteinization were carried out using 1 M hydrochloric acid at 100 °C for 2 h and 1 M NaOH for 4 h at 100 °C, respectively. Biological chitin extraction was carried out by protease-producing bacteria and lactic-acid-producing bacteria for protein and mineral removal, respectively. The extracted chitin was converted to chitosan via deacetylation using 40% NaOH for 8 h at 100 °C. Chitin characterization was done using FTIR spectroscopy, while the antimicrobial properties were determined using the disc diffusion method. Chemical and biological extraction gave a chitin yield of 10.18% and 11.85%, respectively. A maximum chitosan yield of 6.58% was achieved via chemical treatment. From the FTIR results, biological and chemical chitin showed characteristic chitin peaks at 1650 and 1550 cm−1—wavenumbers corresponding to amide I stretching and amide II bending, respectively. There was significant growth inhibition for Escherichia coli, Bacillus subtilis,Pseudomonas aeruginosa,Staphylococcus aureus, and Candida albicans when subjected to 2.5 and 5% concentrations of chitosan. Our findings demonstrate that chitosan from BSF pupal exuviae could be a promising and novel therapeutic agent for drug development against resistant strains of bacteria.

Multipotential Alkaline Protease From a Novel Pyxidicoccus sp. 252: Ecofriendly Replacement to Various Chemical Processes

A newly isolated alkaline protease-producing myxobacterium was isolated from soil. The strain was identified as Pyxidicoccus sp. S252 on the basis of 16S rRNA sequence analysis. The extracellular alkaline proteases produced by isolate S252 (PyCP) was optimally active in the pH range of 11.0–12.0 and temperature range of 40–50°C The zymogram of PyCP showed six caseinolytic protease bands. The proteases were stable in the pH range of 8.0–10.0 and temperature range of 40–50°C. The activity of PyCP was enhanced in the presence of Na+, Mg2+, Cu2+, Tween-20, and hydrogen peroxide (H2O2) (hydrogen peroxide), whereas in Triton X-100, glycerol, ethylenediaminetetraacetic acid (EDTA), and Co2+, it was stable. PyCP showed a potential in various applications. The addition of PyCP in the commercial detergent enhanced the wash performance of the detergent by efficiently removing the stains of tomato ketchup and coffee. PyCP efficiently hydrolyzed the gelatin layer on X-ray film to release the embedded silver. PyCP also showed potent dehairing of goat skin and also efficiently deproteinized sea shell waste indicating its application in chitin extraction. Thus, the results of the present study indicate that Pyxidicoccus sp. S252 proteases have the potential to be used as an ecofriendly replacement of chemicals in several industrial processes.

Recovery of glucosamine and optimization of chitin extraction with acid and alkali process from shells of sea mussel Mytilus galloprovincialis in Greece

Mussel wastes include byproducts such as seed, barnacles fouling, broken shells, byssus threads, which are associated with serious environmental impacts on aquatic ecosystems. The objective of reducing wastes and to avoid environmental problems can be achieved by establishing alternative methods to valorize mussel wastes converting them into high value-added products. The present study aims to obtain by chemical methods the extraction of chitin and glucosamine from shells of Mytilus galloprovincialis. Chitin, which chemically is a linear polysaccharide of β (1→4) linked N-acetylglucosamine monomers, possesses multifunctional properties and is suitable for various applications mainly in pharmaceutical, biomedical food and packaging fields. Glucosamine (C6H13NO5) is an amino sugar (2-amino-2-deoxy-β-D-glucopyranose) and is a part of the structure of two polysaccharides, chitosan and chitin. Due to its high aqueous solubility and physical stability is used as a potential solid-dispersion carrier to improve the biopharmaceutical properties of drugs. The influence of temperature, solid/liquid ratio (g/ml), molarity of solutions and agitation on chitin production and acid hydrolysis of chitin was studied to achieve optimum output of chitin and glucosamine. Our results demonstrates that the production of glucosamine and chitin is greatly influenced by the experimental conditions. The best yield for glucosamine was obtained at a solid/liquid ratio of 1:15 at 60°C for 1 hour.

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.

Chicken Bone as Chitin Source

Chitin extraction has been the interest of many researchers using it as biopolymer in many applications. Many approaches have been adopted in chitin extraction. The aim of this work isto prepare chitin from chicken bone, proposing a new approach, using all chicken bone as source of chitin and recycling this waste instead of accumulation as refuse. A chemical method was used in extractingchitin (deproteinization and demineralization).The obtained chitin was characterized byFourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD), X-ray fluorescence(XRF).The best result obtained was in experiment E5,which are close to the standard results, but chitin extraction was 16.25% of the total weight of the chicken bone used in the experiment. Which is considered a high percentage compared to the state of research on chitin extraction from chicken feat.