Synthetic orange pigment can be substituted by natural carotenoids for the development of different eco-friendly products for biomedical, agricultural, food, and cosmetics industries (Lorenz and Cysewski, 2000; Guerin et al., 2003; Sachindra et al., 2007; Dhankhar et al., 2012). Crustacean wastes are observed to be a significant source of carotenoid pigment-protein complex. Extraction of the unstable carotenoid pigment, astaxanthin (3,3-dihydroxy- p.p-carotene-4,4-dione) from crustacean shell is dependent on the stabilized crustacean waste (Armenta-Lopez et al., 2002; Silva et al., 2018). Besides, the nutraceutical importance of astaxanthin pigment makes its importance to recover from shrimp shell waste during chitin recovery (Prameela et al., 2017). Chemical chitin extraction by sodium hydroxide and hydrochloric acid not only causes the loose of chitin quality, environmental hazards, etc., but also degrades its associated commercially important pigment astaxanthin. Therefore, shrimp wastes are exploited for carotenoid extraction by Lactobacillus based fermentation followed by organic solvent-mediated extraction procedure. In such a process, the protein pigment complex was separated by enzymatic treatment and ultra-filtration to remove 265 kDa protein and stable pigment generation for further use (Armenta-Lopez et al., 2002). In another Lactobacillusplantarum mediated chitin recovery process, the bacterial strain is isolated from Solenocera melantlio gut content and used for economic chitin and environmentally safe astaxanthin recovery from shrimp shell wastes (Prameela et al., 2017).

Microorganism Used



Source of Microbes

Waste Used for Fermentation

Type of Fermentation

Efficiency (%) DM DP


Bacillus subtilis









Sini et al., 2007

Pseudomonas aeruginosa F722


Soil sample of Yeosu

Crab shell

Liquid fermentation



Oh et al., 2007



Soil sample


Liquid fermentation

47 ± 1.2

84 ± 1.6

Jo et al., 2008


Bayou La Batre

Shrimp shell





Sorokulova et al., 2009

Bacillus pumilus A1


Shrimp shell waste

Liquid state



Ghorbel- Bellaaj et al.. 2012

Senatia marcescens











Sedaghat et al., 2016

Bacillus subtilis



Shrimp shell waste

Liquid fermentation



Gamal et al., 2016

Bacterial enzyme alcalase also plays a significant role to extract shrimp waste-based carotenoid protein at around 0.4% concentration of the reaction mixture within 4 li of reaction. In the same process, use of higher enzyme concentration (0.5%) application at 50°C is also leading to good quality protein isolates having antioxidant property for food industry-related applications (Sowmya et ah, 2014).

In a very recent study, spouted bed drying technique is used for powder chitin recovery from shrimp wastes; where, high-quality astaxanthin extraction is standardized by vegetable oil instead of organic solvent-based extraction procedure. In this teclmique, different air temperature and spouting velocities are used to diy the shrimp paste. The obtained product confirms the reproducibility of the process for chitin recoveiy with subsequent palmolein based astaxanthin extraction. The process achieves to produce around 30 pg of astaxanthin from per gram of dried waste sample (Silva et ah, 2018).


Combinations of biological and chemical methods have been also developed by some of researchers. Crude protease from Bacillus cereus SV1, Bacillus licheniformis NH1, Bacillus subtilis A 26 have been used for DP of shrimp shell; where, DM steps were performed by 1.5 mol/L hydrochloric acid to achieve waste-based chitin isolation (Manni et ah, 2010; Younes et ah, 2012). Recently, sea water-mediated chitin isolation from shrimp shell added a new approach in this process development. In this process, shrimp shell fermentation was carried out by shrimp shell fermentation in seawater medium with protease producing bacteria, Bacillus subtilis (В 1), and Bacillus licheniformis (B2). The use of sea water-based medium enhances proteolytic activity of the bacterial strains results in more than 70% DP of waste samples. The substitution of freshwater by seawater in the fermentation medium enhances the DM of the waste up to 94%. This process may be useful for not only the green method for the industrial chitin extraction step but also for conserving freshwater for washing in a different step of chitin extraction (Pachapur et ah, 2016).


Eco-friendly and rapid chitin isolation process development is also revealed by the use of some other biochemical like deep eutectic solvent use. Choline chloride thiourea, choline chloride glycerol choline chloride urea, choline chloride with malic acid is being used for these processes (Zhu et al., 2017; Huang et al., 2018). Previously, a sonication based chitin extraction from shrimp waste of North Atlantic has been carried out along with the use of chemicals like strong mineral acid and bases (Kjartansson et al., 2006). Among different chemical-based chitin isolation processes, choline chloride malonic acid-based eutectic solvent is observed to achieve around 25% chitin isolation yield from lobster shells; where, the chemical prepared chitin yield is observed to be around 20%. Although, isolated chitin from this process is a mixture of two different crystalline powders with different thermal stability. However, porous lobster isolated chitin by chloride malonic acid-based eutectic solvent is thought to be an important biomaterial for biomedical applications and adsoiption based applications (Zhu et al., 2017). In another study, the eutectic non-toxic solvent of natural metabolites has also been used for the rapid and green chitin isolation process by the use of choline chloride and malic acid in combination. The use of the solvent leads to efficient DM and DP which have been revealed by several biochemical and biophysical studies through FTIR (Fourier-transfonn infrared spectroscopy), x-ray diffraction (XRD), thermogravimetric analysis (TGA) and surface electron microscopy (SEM) (Huang et al., 2018). In the same process microwave irradiation with the before mentioned solvent use is observed to improve removal of protein and minerals from shrimp shells. Elimination of environmentally harsh chemicals makes the process eco-friendly for crustacean waste-based chitin isolation (Huang et al., 2018).

Besides the use of eutectic solvent, the use of commercial proteinases plays a significant role in chitin isolation. A comparison of eleven commercial protease uses for chitin isolation in acidic pH leads to effective protein removal from the waste samples. Additionally, formic acid application in the same process improves the good quality chitin. Other than pepsin, all ten protease show the same type of amino acid profiling; where, glycine is observed to be significant amino acid obtained during chitin isolation process (Baron et al., 2017). In another study on crayfish shell waste powder (CSP) based chitin isolation process, Bacillus coagulans LA 204 with proteinase К is used to remove calcium ion along with other proteinaceous material. A 48 h fermentation process with simultaneous enzymatic hydrolysis in a 5 L Bioreactor with unsterilized condition yield more than 90% chitin recovery from the wastes through efficient DP and DM. Here, glucose addition enhances the calcium removal process; where, more than 90% of removed protein is hydrolyzed (Dun et al., 2019). This simultaneous strategy may be a significant future approach for DM and DP of different crustacean waste- based chitin isolation process development.

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