RESUMO
The mycotoxin Ochratoxin A (OTA) causes serious health risks and is found in food products throughout the world. The most promising method to detoxify this compound is biodegradation. In this study, Aspergillus oryzae strain M30011 was isolated and characterized based on its considerable capacity to degrade OTA. The degradation product (compound I) of A. oryzae-treated OTA was isolated, and its toxicity response was also evaluated. Furthermore, the relationships between three key cultivation condition factors affecting the OTA degradation rate were examined using the response surface methodology (RSM). Compound I was identified as ochratoxin α (C11H9O5Cl), and the toxicity response experiments indicated that A. oryzae detoxified OTA to a great extent. A maximum degradation rate of 94% was observed after 72h. This study demonstrates the potential for using A. oryzae to detoxify OTA and suggests that it could be applied in the food industry to improve food safety and quality.
Assuntos
Aspergillus oryzae , Micotoxinas , Ocratoxinas , Biodegradação Ambiental , Contaminação de AlimentosRESUMO
The preparation of oligosaccharides via xylan hydrolysis is an effective way to add value to hemicellulosic material of agricultural waste. The bacterial strain Streptomyces L10608, isolated from soil, contains genes encoding xylanases of glucoside hydrolase family 10/11 (GH10/11), and these have been cloned to catalyze the production of xylooligosaccharide (XOS). To improve the XOS proportion of hydrolysates produced by xylanase, four amino acid residues were substituted by site-directed mutagenesis, and the mutant genes were overexpressed in Escherichia coli. Mutations replaced the codons encoding Asn214 (+2) and Asn86 (-2) by Ala and removed the Ricin B-lectin domain in GH10-xyn, and mutants Y115A (-2) and Y123A (-2) were produced for GH11-xyn. Interestingly, GH10-N86Q had significantly increased hydrolysis of XOS and almost eliminated xylose (X1) to <2.5%, indicating that the -2 binding site of GH10-xyn of L10608 is required for binding with xylotriose (X3). The hydrolytic activity of GH10-N86Q was increased approximately 1.25-fold using beechwood xylan as a substrate and had high affinity for the substrate with a low Km of about 1.85 mg·mL-1. Otherwise, there were no significant differences in enzymatic properties between GH10-N86Q and GH10-xyn. These mutants offer great potential for modification of xylanase with desired XOS hydrolysis.
Assuntos
Substituição de Aminoácidos , Proteínas de Bactérias/metabolismo , Streptomyces/enzimologia , Xilosidases/metabolismo , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Sítios de Ligação , Glucuronatos/metabolismo , Hidrólise , Oligossacarídeos/metabolismo , Ligação Proteica , Streptomyces/genética , Especificidade por Substrato , Xilanos/metabolismo , Xilose/metabolismo , Xilosidases/química , Xilosidases/genéticaRESUMO
Dry-cured meat products are gaining attention owing to their distinctive sensory characteristics and health benefits. In this study, two Debaryomyces hansenii strains were investigated for their potential as starter cultures for dry-cured pork belly products. After preliminary screening, these D. hansenii strains, namely, S20 and S26, both exhibiting with excellent aroma-producing capacity in a dry-cured meat model, were selected as single-strain starter cultures. For comparison, a non-inoculated control was also evaluated. In S20- and S26-inoculated pork belly, yeast dominated the microbiota and improved microbiological safety by suppressing Enterobacteriaceae growth. Compared with the non-inoculated control, the inoculated pork belly yielded higher hardness and redness (a*) values. Starter culture inoculation accelerated proteolysis in pork belly, improving the content of total free amino acids (TFFAs) and several essential free amino acids (Thr, Val, Met, Ile, Leu, and Phe) at the end of processing. Moreover, the inoculated samples exhibited higher levels of fat oxidation-derived aldehydes as well as esters, acids, alcohols and other compounds than the non-inoculated control at the end of the 95-day ripening period. Overall, these findings provide new insights into the application of D. hansenii isolated from dry-cured ham to dry-cured pork belly.
Assuntos
Debaryomyces , Microbiologia de Alimentos , Produtos da Carne , Animais , Produtos da Carne/microbiologia , Produtos da Carne/análise , Suínos , Humanos , Paladar , Valor Nutritivo , Aminoácidos/análise , Manipulação de Alimentos/métodos , Fermentação , Carne de Porco/microbiologia , Carne de Porco/análise , Odorantes/análise , Proteólise , MasculinoRESUMO
An efficient and reliable duplex SYBR Green real-time quantitative PCR (qPCR) method for beef products adulteration detection was developed based on bovine specific and vertebrate universal primers. By analyzing the numbers, positions (Tm value) of melting curve peaks of the duplex PCR products, we simultaneously identified bovine and preliminary screened non-bovine in samples, and also semi-quantified the bovine percentage according to the area ratios of peaks. All of these were necessary for adulteration determination. The specific and universal primers were designed based on mitochondrial genes ND4 and 16S rRNA respectively, their amplicons Tm values were 72.6 ± 0.5 °C and 79-81 °C. There might be some other peaks at 74-78 °C and above 81 °C if non-bovine components existed. Thelimit of detectionwas 1 pgforbovineDNA, and1 - 30 pg fornon-bovineDNAbasedon differentspecies.
Assuntos
Compostos Orgânicos/metabolismo , Reação em Cadeia da Polimerase em Tempo Real/métodos , Carne Vermelha/análise , Animais , Benzotiazóis , Bovinos , Primers do DNA/genética , Diaminas , Qualidade dos Alimentos , QuinolinasRESUMO
A simple and rapid method for animal species identification to prevent food adulteration based on mitochondrial DNA using two independent multiplex polymerase chain reactions (PCRs) and microchip electrophoresis was developed. This method was designed to identify fourteen domestic animals (Group I: cattle, donkey, dog, fox, raccoon-dog, deer and horse; Group II: pig, sheep, goat, chicken, duck, cat and mouse) simultaneously using ten pairs of primers and three of which were degenerate primers. Sequences for species-specific primers were generated based on mitochondrial genes, including 12S rRNA, 16S rRNA, ND2 and CO I. This method was validated in terms of the specificity, sensitivity and practicability, and the developed multiplex PCR method was able to correctly identify animal species of raw meats and processed meat products. The detection limits of two multiplex PCRs were 0.02â¯ng DNA for animal species in Group I and 0.2â¯ng DNA for Group II, respectively.