Development of a Microbial-Assisted Process for Enhanced Astaxanthin Recovery from Crab Exoskeleton Waste

Abstract

Astaxanthin is a xanthophyll carotenoid possessing impressive nutraceutical, antioxidant, and bioactive merits. Traditionally, astaxanthin is extracted from crustacean wastes via solvent extraction methods. However, the rigid structure of shells that comprise complex proteins and chitin challenges the extraction process. This investigation addressed an efficient microbial-assisted method to facilitate astaxanthin recovery from crab exoskeleton waste utilizing chitinolytic and proteolytic microorganisms. Herein, we evaluated the effect of pretreatment of the exoskeleton waste with a newly isolated probiotic strain, Bacillus amyloliquefaciens CPFD8, showing remarkable protease and chitinase activity and a proteolytic Saccharomyces cerevisiae 006-001 before solvent extraction, using acetone/hexane, on astaxanthin recovery. Furthermore, the antioxidant and anti-inflammatory activities of the recovered astaxanthin were inspected. Results revealed that both strains boosted the astaxanthin yield from the crab (Callinectes sapidus) exoskeleton compared with solvent extraction using acetone/hexane. Under optimum conditions, astaxanthin yield was 217 and 91 µg/g crab exoskeleton in samples treated with B. amyloliquefaciens CPFD8 and S. cerevisiae 006-001, respectively. Interestingly, pretreatment of crab exoskeleton waste with B. amyloliquefaciens CPFD8 yielded more than 6-fold astaxanthin compared with the solvent extraction method that yielded just 35 µg/g. This increase could be attributed to the proteolytic activity of B. amyloliquefaciens CPFD8 that rendered deproteinized shell chitin accessible to chitinase, facilitating the penetration of solvents and the recovery of astaxanthin. The recovered astaxanthin exhibited excellent antioxidant activity in scavenging DPPH or ABTS free radicals with IC50 values of 50.93 and 17.56 µg/mL, respectively. In addition, the recovered astaxanthin showed a remarkable anti-inflammatory impact on LPS-induced murine macrophage RAW264.7 cells and significantly inhibited the production of nitric oxide, TNF-α, and IL-6 compared with the untreated control. These findings suggest the potential use of the developed microbial-assisted method utilizing chitinolytic and proteolytic B. amyloliquefaciens CPFD8 to maximize the recovery of bioactive astaxanthin from crab (C. sapidus) exoskeleton waste.

Extraction and Purification of Highly Active Astaxanthin from Corynebacterium glutamicum Fermentation Broth

“…EC 50 values of the positive controls ascorbic acid (22.9 µg/mL) and BHT (22.4 µg/mL) were in line with the published data obtained by Chintong et al [18]. The astaxanthin containing oleoresin of C. glutamicum showed a higher antioxidant activity (EC 50 = 3.7 µg/mL) compared with different extracts from shrimp (17.5 µg/mL; 6.3 µg/mL) [18,55], crab (50.93 µg/mL) [19], Chromochloris zofingiensis (formerly Chlorella zofingiensis) (Chlorophyta) (1040 µg/mL) [56] and the common astaxanthin production host X. dendrorhous (31.79 µg/mL) [45]. We also measured the activity of esterified astaxanthin from H. lacustris (3.2 µg/mL), which was comparable to our corynebacterial extract.…”

“…Common hosts for the production of natural astaxanthin are the microalgae Haematococcus lacustris (formerly Haematococcus pluvialis) (Chlorophyta), the yeast Xanthophyllomyces dendrorhous and the Gram-negative bacterium Paracoccus carotinifaciens [9,12,16]. Another source of astaxanthin that has been exploited are shells of crustaceans that occur as byproducts in food processing [17][18][19].…”

Characterization of a Metallic-Ions-Independent L-Arabinose Isomerase from Endophytic Bacillus amyloliquefaciens for Production of D-Tagatose as a Functional Sweetener

“…It has been believed that enzymatic activity and bioconversion efficacy are directly influenced by the reaction conditions such as temperature and pH [66][67][68]. The purified BAAI was functional and exhibited maximum activity at 45 • C. This operating temperature is slightly greater than that of L-AI derived from Pseudoalteromonas haloplanktis that exhibited maximum activity at 40 • C [45] but lower than that of Bifidobacterium longum NRRL B-41409 [69].…”

Peningkatan stabilitas oksidatif minyak mata tuna dengan metode purifikasi dan penambahan natural astaxanthin (NAst)

“…Nilai IC 50 astaxanthin dari ekstrak kulit udang (L.vannamei) 14,14 ppm dan termasuk kategori antioksidan sangat kuat. Hasil ini menunjukkan bahwa aktivitas penghambatan ABTS astaxanthin kulit udang lebih unggul dibandingkan dengan astaxanthin eksoskelaton kepiting (C. sapidus) dengan nilai IC 50 17,56 dan asam askorbat 28,46 µg/mL (Abd El-Ghany et al, 2023).…”