Abstract

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.

Highlights

  • Chitin is one of the most abundant biopolymers in the world, and these natural polymers have versatile properties, such as biocompatibility and non-toxicity

  • Over 350 bacterial strains isolated from the soils of Northern Taiwan were cultivated at 37 ◦ C in a medium containing 1% squid pen powder (SPP)

  • The TKU036 strain was identified as Paenibacillus sp

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Summary

Introduction

Chitin is one of the most abundant biopolymers in the world, and these natural polymers have versatile properties, such as biocompatibility and non-toxicity. Chitin is obtained from shrimp shells, crab shells, and squid pens using a strong alkali or an inorganic acid for deproteinization or demineralization, respectively [1]. These chemical processes have several drawbacks, such as the creation of pollutant alkali or acid liquid. For recycling squid pens in order to produce additional highly value-added products other than chitin or chitosan, we investigated the reutilization of this fishery processings via microbial conversion in order to produce enzymes [2,3,4,5], exopolysaccharides [6,7], chitooligomers [3], antioxidants [8,9], insecticidal materials [10], and biosorbents [11,12]. The antioxidant and anti-inflammatory activities of the isolated HGA were investigated and compared with those activities of other well-known antioxidant (α–tocopherol) and anti-inflammatory compound (quercetin)

Screening and Identification of Strain TKU036
Culture Conditions for Antioxidant Production
Isolation of Antioxidant Compounds ethanol extract was separated
Identification of HGA and Tryptophan by NMR
The Effect of HGA on Cytotoxicity and Anti-Inflammation
Confirmation of HGA and Tryptophan Produced from SPP by Fermentation
Materials
Antioxidant Activity Assay
Screening of Antioxidant-Producing Strain
Extraction and Isolation of HGA and Tryptophan
The Analysis of HGA and Tryptophan by HPLC
Conclusions
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