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1.
Arch Microbiol ; 205(8): 287, 2023 Jul 16.
Artigo em Inglês | MEDLINE | ID: mdl-37454356

RESUMO

Laccases or laccase-like multicopper oxidases have great potential in bioremediation to oxidase phenolic or non-phenolic substrates. However, their inability to maintain stability in harsh environmental conditions and against non-substrate compounds is one of the main reasons for their limited use. The gene (mco) encoding multicopper oxidase from Bacillus mojavensis TH309 were cloned into pET14b( +), expressed in Escherichia coli, and purified as histidine tagged enzyme (BmLMCO). The molecular weight of the enzyme was about 60 kDa. The enzyme exhibited laccase-like activity toward 2,6-dimethoxyphenol (2,6-DMP), syringaldazine (SGZ), and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS). The highest enzyme activity was recorded at 80 °C and pH 8. BmLMCO showed a half-life of ~ 305, 99, 50, 46, 36, and 20 min at 40, 50, 60, 70, 80, and 90 °C, respectively. It retained more than 60% of its activity after pre-incubation in the range of pH 5-12 for 60 min. The enzyme activity significantly increased in the presence of 1 mM of Cu2+. Moreover, BmLMCO tolerated various chemicals and showed excellent compatibility with organic solvents. The Michaelis constant (Km) and the maximum velocity (Vmax) values of BmLMCO were 0.98 mM and 93.45 µmol/min, respectively, with 2,6-DMP as the substrate. BmLMCO reduced the antibacterial activity of cefprozil, gentamycin, and erythromycin by 72.3 ± 1.5%, 79.6 ± 6.4%, and 19.7 ± 4.1%, respectively. This is the first revealing shows the recombinant production of laccase-like multicopper oxidase from any B. mojavensis strains, its biochemical properties, and potential for use in bioremediation.


Assuntos
Antibacterianos , Lacase , Lacase/genética , Lacase/metabolismo , Antibacterianos/farmacologia , Clonagem Molecular , Especificidade por Substrato , Concentração de Íons de Hidrogênio , Estabilidade Enzimática
2.
Plants (Basel) ; 12(6)2023 Mar 21.
Artigo em Inglês | MEDLINE | ID: mdl-36987087

RESUMO

In the era of global warming, heat stress, particularly at the seedling stage, is a major problem that affects the production and productivity of crops such as mustard that are grown in cooler climates. Nineteen mustard cultivars were exposed to contrasting temperature regimes-20 °C, 30 °C, 40 °C and a variable range of 25-40 °C-and evaluated for changes in physiological and biochemical parameters at the seedling stage to study their role in heat-stress tolerance. Exposure to heat stress showed detrimental effects on seedling growth as revealed by reduced vigor indices, survival percentages, antioxidant activity and proline content. The cultivars were grouped into tolerant, moderately tolerant and susceptible based on the survival percentage and biochemical parameters. All the conventional and three single-zero cultivars were found to be tolerant and moderately tolerant, respectively, while double-zero cultivars were reckoned to be susceptible except for two cultivars. Significant increases in proline content and catalase and peroxidase activities were found associated with thermo-tolerant cultivars. More efficient antioxidant system activity and proline accumulation were noticed in conventional along with three single-zero (PM-21, PM-22, PM-30) and two double-zero (JC-21, JC-33) cultivars that might have provided better protection to them under heat stress than the remaining one single- and nine double-zero cultivars. Tolerant cultivars also resulted in significantly higher values of most of the yield attributing traits. Heat-stress-tolerant cultivars could easily be selected based on the survival percentage, proline and antioxidants at the seedling stage and included as efficient cultivars in breeding programs.

3.
BMC Microbiol ; 23(1): 56, 2023 03 03.
Artigo em Inglês | MEDLINE | ID: mdl-36869305

RESUMO

BACKGROUND: Thermostable microorganisms are extremophiles. They have a special genetic background and metabolic pathway and can produce a variety of enzymes and other active substances with special functions. Most thermo-tolerant microorganisms from environmental samples have resisted cultivation on artificial growth media. Therefore, it is of great significance to isolate more thermo-tolerant microorganisms and study their characteristics to explore the origin of life and exploit more thermo-tolerant enzymes. Tengchong hot spring in Yunnan contains a lot of thermo-tolerant microbial resources because of its perennial high temperature. The ichip method was developed by D. Nichols in 2010 and can be used to isolate so-called "uncultivable" microorganisms from different environments. Here, we describe the first application of modified ichip to isolate thermo-tolerant bacteria from hot springs. RESULTS: In this study, 133 strains of bacteria belonging to 19 genera were obtained. 107 strains of bacteria in 17 genera were isolated by modified ichip, and 26 strains of bacteria in 6 genera were isolated by direct plating methods. 25 strains are previously uncultured, 20 of which can only be cultivated after being domesticated by ichip. Two strains of previously unculturable Lysobacter sp., which can withstand 85 °C, were isolated for the first time. Alkalihalobacillus, Lysobacter and Agromyces genera were first found to have 85 °C tolerance. CONCLUSION: Our results indicate that the modified ichip approach can be successfully applied in a hot spring environment.


Assuntos
Actinomycetales , Bacillaceae , Fontes Termais , China , Meios de Cultura
4.
Microorganisms ; 10(9)2022 Aug 29.
Artigo em Inglês | MEDLINE | ID: mdl-36144343

RESUMO

The production of vinegar on an industrial scale from different raw materials is subject to constraints, notably the low tolerance of acetic acid bacteria (AAB) to high temperatures and high ethanol concentrations. In this study, we used 25 samples of different fruits from seven Moroccan biotopes with arid and semi-arid environmental conditions as a basic substrate to isolate thermo- and ethanol-tolerant AAB strains. The isolation and morphological, biochemical and metabolic characterization of these bacteria allowed us to isolate a total number of 400 strains with characters similar to AAB, of which six strains (FAGD1, FAGD10, FAGD18 and GCM2, GCM4, GCM15) were found to be mobile and immobile Gram-negative bacteria with ellipsoidal rod-shaped colonies that clustered in pairs and in isolated chains. These strains are capable of producing acetic acid from ethanol, growing on peptone and oxidizing acetate to CO2 and H2O. Strains FAGD1, FAGD10 and FAGD18 show negative growth on YPG medium containing D-glucose > 30%, while strains GCM2, GCM4 and GCM15 show positive growth. These six strains stand out on CARR indicator medium as isolates of the genus Acetobacter ssp. Analysis of 16S rDNA gene sequencing allowed us to differentiate these strains as Acetobacter fabarum and Acetobacter pasteurianus. The study of the tolerance of these six isolates towards pH showed that most of the six strains are unable to grow at pH 3 and pH 9, with an ideal pH of 5. The behavior of the six strains at different concentrations of ethanol shows an optimal production of acetic acid after incubation at concentrations between 6% and 8% (v/v) of ethanol. All six strains tolerated an ethanol concentration of 16% (v/v). The resistance of the strains to acetic acid differs between the species of AAB. The optimum acetic acid production is obtained at a concentration of 1% (v/v) for the strains of FAGD1, FAGD10 and FAGD18, and 3% (v/v) for GCM2, GCM4 and GCM15. These strains are able to tolerate an acetic acid concentration of up to 6% (v/v). The production kinetics of the six strains show the highest levels of growth and acetic acid production at 30 °C. This rate of growth and acetic acid production is high at 35 °C, 37 °C and 40 °C. Above 40 °C, the production of acid is reduced. All six strains continue to produce acetic acid, even at high temperatures up to 48 °C. These strains can be used in the vinegar production industry to minimize the load on cooling systems, especially in countries with high summer temperatures.

5.
AMB Express ; 10(1): 44, 2020 Mar 10.
Artigo em Inglês | MEDLINE | ID: mdl-32157462

RESUMO

Two novel glycoside hydrolases were cloned from the genomic DNA of alklinphilic bacterium Cellulomonas bogoriensis 69B4T and functionally expressed in Escherichia coli. The two enzymes shared less than 73% of identities with other known glycosidases and belonged to glycoside hydrolase families 5 and 9. Recombinant Cel5A exhibited optimum activity at pH 5.0 and at a temperature of 70 °C, and Cel9A showed optimum activity at pH 7.0 and at a temperature of 60 °C. The two enzymes exhibited activity at alkaline pH 11 and were stable over a wide range of pH. The maximum activities of Cel5A and Cel9A were observed in 0.5 M NaCl and 1 M KCl, respectively. In addition, these two enzymes exhibited excellent halostability with residual activities of more than 70% after pre-incubation for 6 days in 5 M NaCl or 4 M KCl. Substrate specificity analysis revealed that Cel5A and Cel9A specifically cleaved the ß-1,4-glycosidic linkage in cellulose with the highest activity on carboxymethyl cellulose sodium (78.3 and 145.3 U/mg, respectively). Cel5A is an endoglucanase, whereas Cel9A exhibits endo and exo activities. As alkali-activated, thermo-tolerant, and salt-tolerant cellulases, Cel5A and Cel9A are promising candidates for further research and industrial applications.

6.
Mar Drugs ; 17(5)2019 May 24.
Artigo em Inglês | MEDLINE | ID: mdl-31137685

RESUMO

Alginate oligosaccharides (AOS) show versatile bioactivities. Although various alginate lyases have been characterized, enzymes with special characteristics are still rare. In this study, a polysaccharide lyase family 7 (PL7) alginate lyase-encoding gene, aly08, was cloned from the marine bacterium Vibrio sp. SY01 and expressed in Escherichia coli. The purified alginate lyase Aly08, with a molecular weight of 35 kDa, showed a specific activity of 841 U/mg at its optimal pH (pH 8.35) and temperature (45 °C). Aly08 showed good pH-stability, as it remained more than 80% of its initial activity in a wide pH range (4.0-10.0). Aly08 was also a thermo-tolerant enzyme that recovered 70.8% of its initial activity following heat shock treatment for 5 min. This study also demonstrated that Aly08 is a polyG-preferred enzyme. Furthermore, Aly08 degraded alginates into disaccharides and trisaccharides in an endo-manner. Its thermo-tolerance and pH-stable properties make Aly08 a good candidate for further applications.


Assuntos
Organismos Aquáticos/enzimologia , Polissacarídeo-Liases/metabolismo , Temperatura , Vibrio/enzimologia , Organismos Aquáticos/genética , Estabilidade Enzimática , Escherichia coli/genética , Concentração de Íons de Hidrogênio , Polissacarídeo-Liases/química , Polissacarídeo-Liases/genética , Polissacarídeo-Liases/isolamento & purificação , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Vibrio/genética
7.
Waste Manag ; 68: 64-73, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28647221

RESUMO

The inoculum containing four cellulolytic thermophilic actinomycetes was screened from compost samples, and was inoculated into co-composting during different inoculation phases. The effect of different inoculation phases on cellulose degradation, humic substances formation and the relationship between inoculation and physical-chemical parameters was determined. The results revealed that inoculation at different phases of composting improved cellulase activities, accelerated the degradation of cellulose, increased the content of humic substances and influenced the structure of actinomycetic community, but there were significant differences between different inoculation phases. Redundancy analysis showed that the different inoculation phases had different impacts on the relationship between exogenous actinobacteria and physical-chemical parameters. Therefore, based on the promoting effort of inoculation in thermophilic phase of composting for the formation of humic substances, we suggested an optimized inoculation strategy to increase the content of humic substances, alleviate CO2 emission during composting.


Assuntos
Actinobacteria , Celulose , Compostagem , Actinomyces , Substâncias Húmicas , Solo
8.
Int J Biol Macromol ; 102: 1059-1065, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28435055

RESUMO

ι-Carrageenases play a role in marine ι-carrageenan degradation, and their enzymatic hydrolysates are thought to be excellent antioxidants. In this study, we identified a new ι-carrageenase, encoded by cgiF, in psychrophilic bacterium Flavobacterium sp. YS-80-122. The deduced ι-carrageenase, CgiF, belongs to glycoside hydrolase family 82 and shows less than 40% amino acid identity with characterized ι-carrageenases. The activity of recombinant CgiF peaked at 30°C (1,207.8U/mg). Notably, CgiF is a cold-adapted ι-carrageenase, which showed 36.5% and 57% of the maximum activity at 10°C and 15°C, respectively. In addition, it is a thermo-tolerant enzyme that recovered 58.2% of its initial activity after heat shock. Furthermore, although the activity of CgiF was enhanced by NaCl, the enzyme is active in absence of NaCl. This study also shows that CgiF is an endo-type ι-carrageenase that hydrolyzes ß-1,4-linkages of ι-carrageenan, yielding neo-ι-carratetraose as the main product. Its cold-adaptation, thermo-tolerance, NaCl independence and high neo-ι-carratetraose yield make CgiF an excellent candidate for industrial applications in production of ι-carrageen oligosaccharides from seaweed polysaccharides.


Assuntos
Adaptação Fisiológica , Carragenina/metabolismo , Temperatura Baixa , Flavobacterium/enzimologia , Flavobacterium/fisiologia , Glicosídeo Hidrolases/genética , Glicosídeo Hidrolases/metabolismo , Sequência de Aminoácidos , Clonagem Molecular , Flavobacterium/genética , Glicosídeo Hidrolases/química , Glicosídeo Hidrolases/isolamento & purificação , Especificidade por Substrato
9.
Iran J Microbiol ; 9(5): 295-304, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-29296275

RESUMO

BACKGROUND AND OBJECTIVES: Alkaline pH of the soil facilitates the conversion of phosphate present in phosphate fertilizer applied in the field to insoluble phosphate which is not available to plants. Problem of soluble phosphate deficiency arises, primarily due to needless use of phosphate fertilizer. We sought to biofertilizer with the thermo-tolerant phosphate solubilizing actinomycetes consortium that could convert insoluble phosphate to soluble phosphate at wider temperature range. MATERIALS AND METHODS: In the present investigation consortium of five thermo-tolerant phosphate solubilizing actinomycetes was applied for preparation of inoculum to produce multipurpose bio-fertilizer. Phosphates solubilizing thermo-tolerant 32 actinomycetes strains were processed for identification with the use of PIBWIN software and were screened for phosphate solubilizing activity. RESULTS: Amongst these five actinomycetes were selected on the basis of their ability to produce cellulase, chitinase, pectinase, protease, lipase, amylase and phosphate solubilizing enzymes. Ability to produce these enzymes at 28°C and 50°C were examined. Biofertilizer was prepared by using agricultural waste as a raw material. While preparation of bio-fertilizer the pH decreased from 7.5 to 4.3 and temperature increased up to 74°C maximum at the end of 4th week and in subsequent week it started to decline gradually till it reached around 50°C, which was found to be stable up to eighth week. This thermo-tolerant actinomycetes consortium released soluble phosphate of up to 46.7 µg ml-1. CONCLUSION: As the mesophilic organisms die out at high temperature of composting hence thormo-tolerant actinomycetes would be the better substitute for preparation of phosphate solubilizing bio-fertilizer with added potential to degrade complex macromolecules in composting.

10.
Acta sci., Biol. sci ; 38(3): 333-339, jul.-set. 2016. tab, ilus
Artigo em Inglês | LILACS | ID: biblio-827241

RESUMO

Mangroves are classified as permanent preservation areas and regarded as natural nurseries. However, they have suffered several anthropogenic stresses, resulting in their decline. In the light of that, comes the importance of researching their environmental characteristics and revealing possible factors that have led to the degradation of this important ecosystem. The aim of this study was to evaluate the environmental quality of different areas in the mangroves of Ilha do Lameirão Ecological Station through microbiological analyzes of sediment and interstitial water along ten (10) sites, distributed in two areas with different conservation levels (Canal dos Escravos (CE) and Maria Ortiz (MO)) between 2010 and 2012. The microbiological analyzes revealed that MO region, in all seasons of the year, achieved total coliform and thermo -tolerant coliform values above those permitted by the CONAMA Resolution 357/05, fitting the Class 2 conservation standard. The presence of high levels of total and thermo-tolerant coliforms in MO is a strong indicator of impacts originated from the human population and, consequently, the decline of the mangrove itself and the health of human communities surrounding that area.


Os manguezais são classificados como áreas de preservação permanente e considerados como berçários naturais, no entanto sofrem várias tensões antropogênicas, resultando em seu declínio. À luz disso, evidencia-se a importância de pesquisar suas características ambientais e reveladoras de possíveis fatores que levaram à degradação desse importante ecossistema. O objetivo deste estudo é avaliar a qualidade ambiental das diferentes áreas em manguezais da Estação Ecológica Ilha do Lameirão por meio de análises microbiológicas de sedimentos e da água intersticial ao longo de dez locais, distribuídos em duas áreas com diferentes níveis de conservação (Canal dos Escravos (CE) e Maria Ortiz (MO)) entre 2010 e 2012. As análises microbiológicas revelaram que a região MO, em todas as estações do ano, obteve valores de coliformes termo-tolerantes e totais acima do permitido pela resolução CONAMA 357/05, Classe 2. A presença de níveis elevados de coliformes totais e termo-tolerantes em MO é um forte indicador dos impactos originados da população humana e, consequentemente, do declínio do próprio mangue e da saúde das comunidades humanas que cercam a área.


Assuntos
Coliformes , Microbiologia
11.
FEMS Microbiol Lett ; 363(9)2016 05.
Artigo em Inglês | MEDLINE | ID: mdl-27030725

RESUMO

Two new alginate lyase genes, oalY1 and oalY2, have been cloned from the newly isolated marine bacterium Halomonas sp. QY114 and expressed in Escherichia coli The deduced alginate lyases, OalY1 and OalY2, belonged to polysaccharide lyase (PL) family 17 and showed less than 45% amino acid identity with all of the characterized oligoalginate lyases. OalY1 and OalY2 exhibited the highest activities at 45°C and 50°C, respectively. Both of them showed more than 50% of the highest activity at 60°C, and 20% at 80°C. In addition, they were salt-dependent and salt-tolerant since both of them showed the highest activity in the presence of 0.5 M NaCl and preserved 63% and 68% of activity in the presence of 3 M NaCl. Significantly, OalY1 and OalY2 could degrade both polyM and polyG blocks into alginate monosaccharides in an exo-lytic type, indicating that they are bifunctional alginate lyases. In conclusion, our study indicated that OalY1 and OalY2 are good candidates for alginate saccharification application, and the salt-tolerance may present an exciting new concept for biofuel production from native brown seaweeds.


Assuntos
Halomonas/enzimologia , Polissacarídeo-Liases/genética , Polissacarídeo-Liases/metabolismo , Tolerância ao Sal , Termotolerância , Alginatos/metabolismo , Biocombustíveis , Clonagem Molecular , Eletroforese em Gel de Poliacrilamida , Escherichia coli/genética , Ácido Glucurônico/metabolismo , Halomonas/genética , Ácidos Hexurônicos/metabolismo , Concentração de Íons de Hidrogênio , Filogenia , Polissacarídeo-Liases/química , Polissacarídeo-Liases/classificação , Cloreto de Sódio/metabolismo , Especificidade por Substrato
12.
Food Chem ; 199: 516-23, 2016 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-26776003

RESUMO

A xylanase gene of glycoside hydrolase family 10, GtXyn10, was cloned from Gloeophyllum trabeum CBS 900.73 and expressed in Pichia pastoris GS115. Purified recombinant GtXyn10 exhibited significant activities to xylan (100.0%), lichenan (11.2%), glucan (15.2%) and p-nitrophenol-ß-cellobiose (18.6%), demonstrated the maximum xylanase and glucanase activities at pH 4.5-5.0 and 75°C, retained stability over the pH range of 2.0-7.5 and at 70°C, and was resistant to pepsin and trypsin, most metal ions and SDS. Multiple sequence alignment and modeled-structure analysis identified a unique Gly48 in GtXyn10, and site-directed mutagenesis of Gly48 to Lys improved the temperature optimum up to 80°C. Under simulated mashing conditions, GtXyn10 (80U) reduced the mash viscosity by 12.8% and improved the filtration rate by 31.3%. All these properties above make GtXyn10 attractive for potential applications in the feed and brewing industries.


Assuntos
Endo-1,4-beta-Xilanases/química , Clonagem Molecular , Digestão , Fermentação , Microbiologia Industrial , Pichia/genética
13.
Adv Pharm Bull ; 6(4): 551-561, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28101462

RESUMO

Purpose: Robust pharmaceutical and industrial enzymes from extremophile microorganisms are main source of enzymes with tremendous stability under harsh conditions which make them potential tools for commercial and biotechnological applications. Methods: The genome of a Gram-positive halo-thermotolerant Bacillus sp. SL1, new isolate from Saline Lake, was investigated for the presence of genes coding for potentially pharmaceutical enzymes. We determined gene sequences for the enzymes laccase (CotA), l-asparaginase (ansA3, ansA1), glutamate-specific endopeptidase (blaSE), l-arabinose isomerase (araA2), endo-1,4-ß mannosidase (gmuG), glutaminase (glsA), pectate lyase (pelA), cellulase (bglC1), aldehyde dehydrogenase (ycbD) and allantoinases (pucH) in the genome of Bacillus sp. SL1. Results: Based on the DNA sequence alignment results, six of the studied enzymes of Bacillus sp. SL-1 showed 100% similarity at the nucleotide level to the same genes of B. licheniformis 14580 demonstrating extensive organizational relationship between these two strains. Despite high similarities between the B. licheniformis and Bacillus sp. SL-1 genomes, there are minor differences in the sequences of some enzyme. Approximately 30% of the enzyme sequences revealed more than 99% identity with some variations in nucleotides leading to amino acid substitution in protein sequences. Conclusion: Molecular characterization of this new isolate provides useful information regarding evolutionary relationship between B. subtilis and B. licheniformis species. Since, the most industrial processes are often performed in harsh conditions, enzymes from such halo-thermotolerant bacteria may provide economically and industrially appealing biocatalysts to be used under specific physicochemical situations in medical, pharmaceutical, chemical and other industries.

14.
Enzyme Microb Technol ; 70: 72-8, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25659635

RESUMO

In this paper, glutamate dehydrogenase (Gldh) is reported to efficiently display on Escherichia coli cell surface by using N-terminal region of ice the nucleation protein as an anchoring motif. The presence of Gldh was confirmed by SDS-PAGE and enzyme activity assay. Gldh was detected mainly in the outer membrane fraction, suggesting that the Gldh was displayed on the bacterial cell surface. The optimal temperature and pH for the bacteria cell-surface displayed Gldh (bacteria-Gldh) were 70°C and 9.0, respectively. Additionally, the fusion protein retained almost 100% of its initial enzymatic activity after 1 month incubation at 4°C. Transition metal ions could inhibit the enzyme activity to different extents, while common anions had little adverse effect on enzyme activity. Importantly, the displayed Gldh is most specific to l-glutamate reported so far. The bacterial Gldh was enabled to catalyze oxidization of l-glutamate with NADP(+) as cofactor, and the resultant NADPH can be detected spectrometrically at 340nm. The bacterial-Gldh based l-glutamate assay was established, where the absorbance at 340nm increased linearly with the increasing l-glutamate concentration within the range of 10-400µM. Further, the proposed approach was successfully applied to measure l-glutamate in real samples.


Assuntos
Proteínas de Escherichia coli/metabolismo , Escherichia coli/enzimologia , Glutamato Desidrogenase/metabolismo , Ácido Glutâmico/análise , Ânions/farmacologia , Proteínas da Membrana Bacteriana Externa/genética , Proteínas da Membrana Bacteriana Externa/metabolismo , Biotecnologia , Membrana Celular/enzimologia , Estabilidade Enzimática , Enzimas Imobilizadas/genética , Enzimas Imobilizadas/metabolismo , Escherichia coli/genética , Proteínas de Escherichia coli/genética , Glutamato Desidrogenase/genética , Concentração de Íons de Hidrogênio , Metais/farmacologia , Engenharia de Proteínas , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Especificidade por Substrato , Temperatura
15.
Bioresour Technol ; 170: 38-44, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25118151

RESUMO

In this study, thermo-tolerant devices consisting of heat shock genes from thermophiles were designed and introduced into Saccharomyces cerevisiae for improving its thermo-tolerance. Among ten engineered thermo-tolerant yeasts, T.te-TTE2469, T.te-GroS2 and T.te-IbpA displayed over 25% increased cell density and 1.5-4-fold cell viability compared with the control. Physiological characteristics of thermo-tolerant strains revealed that better cell wall integrity, higher trehalose content and enhanced metabolic energy were preserved by thermo-tolerant devices. Engineered thermo-tolerant strain was used to investigate the impact of thermo-tolerant device on pathway efficiency by introducing ß-amyrin synthesis pathway, showed 28.1% increased ß-amyrin titer, 28-35°C broadened growth temperature range and 72h shortened fermentation period. The results indicated that implanting heat shock proteins from thermophiles to S. cerevisiae would be an efficient approach to improve its thermo-tolerance.


Assuntos
Adaptação Fisiológica/genética , Vias Biossintéticas/genética , Engenharia Genética/métodos , Proteínas de Choque Térmico/genética , Temperatura Alta , Saccharomyces cerevisiae/fisiologia , Análise de Variância , Vias Biossintéticas/fisiologia , Sobrevivência Celular/fisiologia , Vermelho Congo , Ácido Oleanólico/análogos & derivados , Ácido Oleanólico/biossíntese , Reação em Cadeia da Polimerase , Saccharomyces cerevisiae/genética , Trealose/metabolismo
16.
Arq. Inst. Biol ; 81(1): 30-35, mar. 2014. tab
Artigo em Português | LILACS, VETINDEX | ID: biblio-909143

RESUMO

O objetivo deste trabalho foi avaliar o perfil das características físico-químicas e microbiológicas dos queijos de coalho comercializados em seis estados do nordeste brasileiro. Foram realizadas análises físico-química e microbiológica de 104 amostras de queijo de coalho comercializadas nos estados de Pernambuco, Piauí, Ceará, Rio Grande do Norte, Sergipe e Paraíba, sendo 50 amostras inspecionadas e 54 sem inspeção. Não foi observada diferença significativa (p > 0,05) para os parâmetros físico-químicos entre as amostras. De acordo com os parâmetros legislativos, nas 104 amostras analisadas de queijos tipo coalho, verificaram-se que 100 (96,15%) estavam fora dos limites para Staphylococcus coagulase positiva; 32 amostras (31%) também não seguiam a padronização exigida para coliformes termotolerantes; e, em apenas uma amostra do total, constatou-se a presença de Salmonella sp. Não foi observada diferença significativa (p > 0,05) entre as amostras com e sem inspeção para as análises microbiológicas. A semelhança encontrada na avaliação dos parâmetros físico-químicos do queijo de coalho entre os estados avaliados demonstra uniformidade no processamento desse produto. Altos níveis de contaminação por agentes patogênicos, como o Staphylococcus coagulase positiva e coliformes termotolerantes, além da presença de Salmonella sp., demonstram que o consumo de queijo de coalho pode representar risco à saúde dos consumidores.(AU)


The objective of this study was to evaluate the physical-chemical and microbiological profile of the rennet cheese sold in six states in northeastern Brazil. Physical-chemical and microbiological analyses of 104 samples of rennet cheese sold in the states of Pernambuco, Piauí, Ceará, Rio Grande do Norte, Paraíba and Sergipe were analyzed, being 50 samples inspected and 54 without inspection. There was no significant difference (p > 0.05) for physical-chemical parameters between samples. In accordance with the parameters of the legislation, in the 104 examined samples of rennet cheese, it was observed that 100 (96.15%) were outside the limits for Staphylococcus coagulase positive; 32 samples (31%) did not follow the standardization required for coliforms; and only one sample showed the presence of Salmonella sp. There was no significant difference (p > 0.05) between samples with and without inspection for microbiological analyses. The similarity found in the assessment of physical-chemical parameters of rennet cheese between the evaluated states demonstrates uniformity in the processing of this product. High levels of contamination by pathogens such as Staphylococcus coagulase positive and thermo-tolerant coliforms, as well as the presence of Salmonella sp., demonstrate that the consumption of rennet cheese may present a health risk for consumers.(AU)


Assuntos
Salmonella/virologia , Staphylococcus/virologia , Contaminação de Alimentos , Queijo/análise , Coliformes , Fenômenos Químicos/métodos , Técnicas Microbiológicas/métodos
17.
Braz. arch. biol. technol ; 52(6): 1363-1371, Nov.-Dec. 2009. graf, tab
Artigo em Inglês | LILACS | ID: lil-539103

RESUMO

Effects of xylose on xylanase production by a thermophilic Bacillus sp showed diverse patterns on corn cob (CC) and wheat bran (WB) as sole carbon sources in solid- state fermentation (SSF) and submerged fermentation (SmF). Supplementation of these media with either mineral salt solution (MSS) or yeast extract peptone (YEP) also exerted variable effects. While under SSF, xylose stimulated xylanase synthesis by 44.01 percent, on wheat bran supplemented with MSS, it decreased the enzyme activity by 12.89 percent with YEP supplementation. In SmF, however the enzyme synthesis was stimulated by xylose on supplementation with both MSS and YEP by 41.38 percent and 27.47 percent, respectively. On corn cob under SSF, xylose repression was significant both with MSS (26.92 percent) and YEP (23.90 percent) supplementation. Repression by xylose also took place on corn cob and YEP (19.69 percent) under SmF, while significant stimulation (28.55 percent) was observed by MSS supplementation. The possible role of media composition and fermentation conditions in the regulation of xylanase synthesis by xylose is discussed.

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