Shrimp Head Powder Research Articles

Utilization of Shrimp Head Wastes into Powder Form as Raw Material for Value-Added Products

Shrimp processing industries produce large amounts of wastes in the form of shrimp heads, shells, and tails, which are more or less 50% of the raw materials. Improper disposal of these nutrient-rich wastes can cause environmental problems if not duly managed. This study aims to utilize Penaeus monodon shrimp head wastes into powder form using a cabinet type drier and establish the processing yield and important product qualities. Two treatments of shrimp head powders were studied: shrimp head powder with carapace (SHPwc) and shrimp head powder without carapace (SHPwoc). The yields obtained were 26.72% for SHPwc while 20.42% for SHPwoc; both are considered to have significant value since both are produced from wastes. Both products have satisfactory water activity levels; however, the bulk density was high, and the solubility was lower than other published powdered seafood by-products. Both products have light orange or brown color: however, in both odor and flavor, SHPwoc had a higher mean general acceptability, with “like slightly-like moderately” results. SHPwoc was subjected to a shelf-life study with added salt and ascorbic acid preservatives. Monthly analyses revealed that the microbiological parameters are still within limits, and both moisture and pH values were acceptable after eight months of storage at ambient temperature (28-30°C). However, the peroxide value is acceptable until only the sixth month, which signifies the end of shelf-life based on theoretical sensory changes. The results of this study show the promising significance of utilizing shrimp processing wastes into seafoodbased products.

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.

Bioproduction of Prodigiosin from Fishery Processing Waste Shrimp Heads and Evaluation of Its Potential Bioactivities

The aim of this work was to reuse a fish processing waste, shrimp head powder (SHP), for the production of prodigiosin (PG) via microbial technology and to assess its potential bioactivities. PG was produced in a 12 L-bioreactor system, and the highest PG productivity of 6310 mg L−1 was achieved when Serratia marcescens CC17 was used for fermentation in a novel designed medium (6.75 L) containing 1.5% C/N source (SHP/casein = 9/1), 0.02% K2SO4, ans 0.025% Ca3(PO4)2, with initial pH 7.0, and fermentation was performed at 28 °C for 8 h. The purified PG showed moderate antioxidants, efficient anti-NO (anti-nitric oxide), and acetylcholinesterase (AChE) inhibitory activities. In a docking study, PG showed better binding energy scores (−12.3 kcal/mol) and more interactions (6 linkages) with several prominent amino acids in the biding sites on AChE that were superior to those of Berberine chloride (−10.8 kcal/mol and one linkage). Notably, this is the first investigation using shrimp heads for the mass bioproduction of PG with high productivity, and Ca3(PO4)2 salt was also newly found to significantly enhance PG production by S. marcescens. This study also provided available data on the anti-NO and anti-AChE effects of PG, especially from the docking simulation PG towards AChE that was described for the first time in this study. The above results suggest that SHP is a good material for the cost-effective bioproduction of PG, which is a potential candidate for anti-NO and anti-Alzheimer drugs.

Microbial Conversion of Shrimp Heads to Proteases and Chitin as an Effective Dye Adsorbent.

As a green and effective technique in the production of a large number of valuable products, the microbial conversion of chitinous fishery wastes is receiving much attention. In this study, protease production using the Paenibacillus mucilaginosus TKU032 strain was conducted on culture media containing several common types of chitinous fishery by-products serving as the carbon and nitrogen (C/N) nutrition source. Among the chitinous wastes, 1.5% (w/v) shrimp head powder (SHP) was found to be the most appropriate nutritional source for protease production when a maximal enzyme activity of 3.14 ± 0.1 U/mL was observed on the 3rd day of the culture period. The molecular mass of P. mucilaginosus TKU032 protease was estimated to be nearly 32 kDa by the polyacrylamide gel electrophoresis method. The residual SHP obtained from the culture medium was also considered to be utilized for chitin extraction. The deproteinization rate of the fermentation was estimated to be 45%, and the chitin obtained from fermented SHP (fSHP) displayed a similar characteristic Fourier-transform infrared spectroscopy (FTIR) profile as that from SHP. In addition, SHP, fSHP, and chitins obtained from SHP and fSHP were investigated for their adsorptive capacity of nine types of dyes, and chitin obtained from fSHP displayed a good adsorption rate on Congo Red and Red No. 7, at 99% and 97%, respectively. In short, the results provide potential support for the utilization of SHP in the production of P. mucilaginosus TKU032 protease via the fermentation as well as the preparation of chitin from fSHP as an effective dye adsorbent.

An Exochitinase with N-Acetyl-β-Glucosaminidase-Like Activity from Shrimp Head Conversion by Streptomyces speibonae and Its Application in Hydrolyzing β-Chitin Powder to Produce N-Acetyl-d-Glucosamine.

Marine chitinous byproducts possess significant applications in many fields. In this research, different kinds of fishery chitin-containing byproducts from shrimp (shrimp head powder (SHP) and demineralized shrimp shell powder), crab (demineralized crab shell powder), as well as squid (squid pen powder) were used to provide both carbon and nitrogen (C/N) nutrients for the production of an exochitinase via Streptomyces speibonae TKU048, a chitinolytic bacterium isolated from Taiwanese soils. S. speibonae TKU048 expressed the highest exochitinase productivity (45.668 U/mL) on 1.5% SHP-containing medium at 37 °C for 2 days. Molecular weight determination analysis basing on polyacrylamide gel electrophoresis revealed the mass of TKU048 exochitinase was approximately 21 kDa. The characterized exochitinase expressed some interesting properties, for example acidic pH optima (pH 3 and pH 5–7) and a higher temperature optimum (60 °C). Furthermore, the main hydrolysis mechanism of TKU048 exochitinase was N-acetyl-β-glucosaminidase-like activity; its most suitable substrate was β-chitin powder. The hydrolysis experiment revealed that TKU048 exochitinase was efficient in the cleavage of β-chitin powder, thereby releasing N-acetyl-d-glucosamine (GlcNAc, monomer unit of chitin structure) as the major product with 0.335 mg/mL of GlcNAc concentration and a yield of 73.64% after 96 h of incubation time. Thus, TKU048 exochitinase may have potential in GlcNAc production due to its N-acetyl-β-glucosaminidase-like activity.

Conversion of Shrimp Head Waste for Production of a Thermotolerant, Detergent-Stable, Alkaline Protease by Paenibacillus sp.

Fishery processing by-products have been of great interest to researchers due to their beneficial applications in many fields. In this study, five types of marine by-products, including demineralized crab shell, demineralized shrimp shell, shrimp head, shrimp shell, and squid pen, provided sources of carbon and nitrogen nutrition by producing a protease from Paenibacillus sp. TKU047. Strain TKU047 demonstrated the highest protease productivity (2.98 U/mL) when cultured for two days on a medium containing 0.5% of shrimp head powder (SHP). The mass of TKU047 protease was determined to be 32 kDa (approximately). TKU047 protease displayed optimal activity at 70–80 °C and pH 9, with a pH range of stability from 6 to 11. TKU047 protease also showed stability in solutions containing surfactants and detergents. Based on its excellent properties, Paenibacillus sp. TKU047 protease may be a feasible candidate for inclusion in laundry detergents.

Anti-α-Glucosidase Activity by a Protease from Bacillus licheniformis.

Anti-α-glucosidase (AAG) compounds have received great attention due to their potential use in treating diabetes. In this study, Bacillus licheniformis TKU004, an isolated bacterial strain from Taiwanese soil, produced AAG activity in the culture supernatant when squid pens were used as the sole carbon/nitrogen (C/N) source. The protein TKU004P, which was isolated from B. licheniformis TKU004, showed stronger AAG activity than acarbose, a commercial anti-diabetic drug (IC50 = 0.1 mg/mL and 2.02 mg/mL, respectively). The molecular weight of TKU004P, determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), was 29 kDa. High-performance liquid chromatography (HPLC) analysis showed that TKU004P may be a protease that demonstrates AAG activity by degrading yeast α-glucosidase. Among the four chitinous sources of C/N, TKU004P produced the highest AAG activity in the culture supernatant when shrimp head powder was used as the sole source (470.66 U/mL). For comparison, 16 proteases, were investigated for AAG activity but TKU004P produced the highest levels. Overall, the findings suggest that TKU004P could have applications in the biochemical and medicinal fields thanks to its ability to control the activity of α-glucosidase.

Production of potent antidiabetic compounds from shrimp head powder via Paenibacillus conversion

Abstract Natural α-glucosidase inhibitors (aGIs) are of great interest as an efficacious and safe therapy for type 2 diabetes, which is an ongoing global health issue. The aim of this study is to utilize shrimp head powder (SHP), an abundant and low-cost material, for the biosynthesis, isolation, and identification of active antidiabetic compounds. SHP was efficiently converted to aGIs via Paenibacillus sp. TKU042 fermentation. Fermented SHP (fSHP) by this strain possesses high pH stability, and stronger yeast α-glucosidase inhibitory activity (92%) than that of acarbose (60%). aGI productivity increased more than 2-fold after optimization (from 225 U/mL to 560 U/mL). Further bioactivity-guided isolation of two major active compounds were identified as nicotinic acid and adenine. Notably, these inhibitors were non-sugar-based moiety aGIs, which is newly isolated and identified from fSHP in this study. In the tests of specific enzyme inhibitory activity, adenine showed highly specific inhibition against yeast α-glucosidase (IC50 = 22 μg/mL); nicotinic acid demonstrated good effect on rat α-glucosidase (IC50 = 70 μg/mL); while acarbose possessed efficient effect on bacteria, rice, and rat α-glucosidases (IC50 = 0.03–108 μg/mL). The current results suggest that it is cost-effective to produce potent aGIs from SHP via Paenibacillus conversion, and these active constituents may be useful in type 2 diabetes management.

Development of Dry Artificial Fish Bait for Trap Fishing Using Tuna Red Meat and Shrimp Head Wastes

ABSTRACT This study developed and evaluated dry artificial fish bait for trap fishing utilizing tuna red meat (TRM) and shrimp head powder (SHP), aiming to replace the usage of edible fishes, such as sardines and squids, as bait. A total of 23 dry baits were prepared and tested for stability in seawater and acceptance by tilapia. Based on the stability and acceptability test results, two baits were selected as ideal and were further tested for protein leaching and dry matter loss in seawater. The tests revealed that the stick-shaped bait containing 61% TRM and 15% SHP was the ideal bait, with a protein leaching rate of 24.82 mg/g/h and dry matter loss of 36.6 mg/g/h. The catch rate of the traps baited with the artificial fish bait was significantly higher (P < 0.05) than those baited with squid meat (17 Nos/trap) and sardines (23 Nos/trap). Further, the catch rate of traps baited with artificial bait (38 Nos/trap) was found to be comparable with those baited with raw shrimp head wastes (SHW) (40 Nos/trap). When the bait was concentrated with 0.1% sodium benzoate, shelf life was extended by up to 6 months.

Reclamation of shrimp heads for the production of α-glucosidase inhibitors by Staphylococcus sp. TKU043

Alpha-glucosidase inhibitors (aGIs) have received much attention due to their important use in treating diabetes mellitus. Although some synthetic aGIs have been available for a long time, they often cause various unexpected side effects. The purpose of this study was to reclaim shrimp heads for the production of aGIs via bacteria. Staphylococcus sp. TKU043, a bacterium strain isolated from Taiwanese soil produced aGIs in a shrimp-head-containing medium. Maximum productivity of aGIs (1457 U/mL) was found after cultivation in 1% shrimp head powder medium for 3 days. The aGI partially purified by column chromatography showed thermal instability, and presumably was a proteinous material. The inhibitor mode of this aGI was determined to be a mixed inhibition mode (non-competitive–uncompetitive inhibition).

Conversion of shrimp heads to α-glucosidase inhibitors via co-culture of Bacillus mycoides TKU040 and Rhizobium sp. TKU041

Alpha-glucosidase inhibitors (aGIs) have potential use as antidiabetic drugs for the treatment of type II diabetes. Most aGIs place a burden on the liver and cause gastrointestinal distress, therefore the development of new aGIs has become very important. In this study, we investigated the production of aGIs by the co-culture of Bacillus mycoides TKU040 and Rhizobium sp. TKU041 using shrimp head powder (SHP) as the sole source of carbon and nitrogen. After fermentation in 50 mL of 1% SHP-containing medium (0.1% K2HPO4 and 0.05% MgSO4·7H2O, pH 9.2) at 37 °C for 4 days, the maximum productivity of aGIs (143 U/mL) was reached. The IC50 of the aGIs produced in the culture supernatant was 3 mg/mL. The aGI activity was only 60% after treatment at pH 3 for 30 min; this increased to 140% after treatment at pH 11 for 30 min. The aGI activity remained at 60% after treatment at 60 °C for 30 min.

Reclamation of Marine Chitinous Materials for the Production of α-Glucosidase Inhibitors via Microbial Conversion.

Six kinds of chitinous materials have been used as sole carbon/nitrogen (C/N) sources for producing α-glucosidase inhibitors (aGI) by Paenibacillus sp. TKU042. The aGI productivity was found to be highest in the culture supernatants using demineralized crab shell powder (deCSP) and demineralized shrimp shell powder (deSSP) as the C/N source. The half maximal inhibitory concentration (IC50) and maximum aGI activity of fermented deCSP (38 µg/mL, 98%), deSSP (108 µg/mL, 89%), squid pen powder (SPP) (422 µg/mL, 98%), and shrimp head powder (SHP) (455 µg/mL, 92%) were compared with those of fermented nutrient broth (FNB) (81 µg/mL, 93%) and acarbose (1095 µg/mL, 74%), a commercial antidiabetic drug. The result of the protein/chitin ratio on aGI production showed that the optimal ratio was 0.2/1. Fermented deCSP showed lower IC50 and higher maximum inhibitory activity than those of acarbose against rat intestinal α-glucosidase.

Conversion of Squid Pen to Homogentisic Acid via Paenibacillus sp. TKU036 and the Antioxidant and Anti-Inflammatory Activities of Homogentisic Acid.

The culture supernatant of Paenibacillus sp. TKU036, a bacterium isolated from Taiwanese soils, showed high antioxidant activity (85%) when cultured in a squid pen powder (SPP)-containing medium at 37 °C for three days. Homogentisic acid (2,5-dihydroxyphenylacetic acid, HGA) was isolated and found to be the major antioxidant in the culture supernatant of the SPP-containing medium fermented by Paenibacillus sp. TKU036. Tryptophan was also present in the culture supernatant. The results of high-performance liquid chromatography (HPLC) fingerprinting showed that HGA and tryptophan were produced via fermentation but did not pre-exist in the unfermented SPP-containing medium. Neither HGA nor tryptophan was found in the culture supernatants obtained from the fermentation of nutrient broth or other chitinous material, i.e., medium containing shrimp head powder, by Paenibacillus sp. TKU036. The production of HGA via microorganisms has rarely been reported. In this study, we found that squid pen was a potential carbon and nitrogen source for Paenibacillus sp. Tryptophan (105 mg/L) and HGA (60 mg/L) were recovered from the culture supernatant. The isolated HGA was found to have higher antioxidant activity (IC50 = 6.9 μg/mL) than α-tocopherol (IC50 = 17.6 μg/mL). The anti-inflammatory activity of the isolated HGA (IC50 = 10.14 μg/mL) was lower than that of quercetin (IC50 = 1.14 μg/mL). As a result, squid pen, a fishery processing byproduct, is a valuable material for the production of tryptophan and the antioxidant and anti-inflammatory HGA via microbial conversion.

Purification of a thermostable chitinase from Bacillus cereus by chitin affinity and its application in microbial community changes in soil

A thermostable chitinase was purified by chitin affinity from the culture supernatant of Bacillus cereus TKU028 with shrimp head powder (SHP) as the sole carbon/nitrogen source. TKU028 chitinase was purified using a one-step affinity adsorbent system, and the molecular mass of TKU028 chitinase (approximately 40 kDa) was then determined using SDS-PAGE. The enzyme was stable for 60 min at temperatures below 60 °C and stable over a broad pH range of 4-9 for 60 min. In addition, the temporal changes of a bacterial community in mangrove river sediment of the Tamsui River with added SHP were also analysed by PCR-denaturing gradient gel electrophoresis to investigate the effects of B. cereus TKU028 on the degradation of SHP. The 6-week incubation sample of SHP and B. cereus TKU028-amended mangrove river sediment displayed the highest amount of biomass, reducing sugar and total sugar, and some variance of bacterial community composition existed in the soils.

Applied development of crude enzyme from Bacillus cereus in prebiotics and microbial community changes in soil

The chitosanase and chitinase activity were revealed in the culture supernatant of Bacillus cereus TKU027 with shrimp head powder (SHP) as the sole carbon/nitrogen source. The chitosan with 60% degree of deacetylation (DD) was depolymerized by TKU027 crude enzyme. The low DP oligomers stimulated the growth of Lactobacillus paracasei BCRC12193 and Lactobacillus kefir BCRC14011 in a MRS broth supplemented with low DP oligomers for 12h. Conversely, the high DP oligomers (0.1%) had potent inhibitory effects against L. paracasei BCRC12193 and L. kefir BCRC14011 for 48h. Besides, the study also investigated the effects of B. cereus TKU027 on degradation of SHP and the survival conditions of bacteria in mangrove river sediment of Tamsui River. The 5 weeks-incubation sample of SHP and B. cereus TKU027-amended mangrove river sediment showed the highest amounts of biomass, reducing sugar and total sugar, and some variance of bacterial community compositions existed in the soils.

Production and purification of a protease, a chitosanase, and chitin oligosaccharides by Bacillus cereus TKU022 fermentation

A protease- and chitosanase-producing strain was isolated and identified as Bacillus cereus TKU022. The protease and chitosanase were both produced using 1.5% (w/v) shrimp head powder (SHP) as the sole carbon/nitrogen source, and these enzymes were purified from the culture supernatant. The molecular masses of the TKU022 protease and chitosanase determined using SDS–PAGE were approximately 45 and 44kDa, respectively. The high stability of the TKU022 protease toward surfactants, an optimal pH of 10 and an optimal temperature of 50–60°C suggest that this high-alkaline protease has potential applications for various industrial processes. Concomitant with the production of the TKU022 chitosanase, N-acetyl chitooligosaccharides were also observed in the culture supernatant, including (GlcNAc)2, (GlcNAc)4, (GlcNAc)5, and (GlcNAc)6 at concentrations of 201.5, 12.4, 0.5, and 0.3μg/mL, respectively, as determined using an HPLC analysis. The chitin oligosaccharides products were also characterized using a MALDI-TOF mass spectrometer. A combination of the HPLC and MALDI-TOF MS results showed that the chitin oligosaccharides of the TKU022 culture supernatant comprise oligomers with degree of polymerization (DP) from 2 to 6. Using this method, the production of a protease, a chitosanase, and chitin oligosaccharides may be useful for various industrial and biological applications.

Fermented and enzymatic production of chitin/chitosan oligosaccharides by extracellular chitinases from Bacillus cereus TKU027

Two chitinases, Chi I and Chi II, were purified from the culture supernatant of Bacillus cereus TKU027 with shrimp head powder (SHP) as the sole carbon/nitrogen source. The molecular masses of Chi I and Chi II determined using SDS–PAGE were approximately 65kDa and 63kDa, respectively. Chi I toward various surfactants showed high stability, such as SDS, Tween 20, Tween 40 and Triton X-100, and these surfactants were stimulator of Chi I chitinase activity. Concomitant with the production of Chi I and Chi II, chitin oligosaccharides were also observed in the culture supernatant, including chitobiose, chitotriose, chitotetrose and chitopentose at concentrations of 0.44mg/mL, 0.08mg/mL, 0.09mg/mL and 0.43mg/mL, respectively. Chitosan with 60% deacetylation was degraded by TKU027 crude enzyme to prepare chitooligosaccharides. MALDI-TOF MS analysis of the enzymatic hydrolyzates indicated that the products were mainly chitooligosaccharides with degree of polymerization (DP) in the 4–9 range.