Factorial design for collagenase production by Penicillium sp. selected from the Caatinga soil

Collagenolytic proteases can hydrolyze both denatured and native collagen and are becoming increasingly important commercially. The aim of this work was to select a new strain of Penicillium sp. isolated from the soil of Caatinga for collagenase production. A factorial design 24 was performed to determine the best conditions of enzyme production. Collagenolytic activity reported on this work is about 2 times larger than existing data. According to the growth kinetics, after 126 hours of production, were obtained the highest values of collagenolytic activity and specific activity. The highest values of collagenolytic activity and specific activity were obtained on a culture medium containing 0.25% (w/v) gelatin, 200 rpm, pH 8.0 and 24 °C. Only two factors were statistically significant: pH and temperature, both with negative effects. The experimental design made possible to define fermentation culture conditions able to increase by 66% the value of the initial enzyme activity.Fundação de Amparo à Ciência e Tecnologia do Estado de Pernambuco (FACEPE) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Effect of Polydextrose on the Growth of Pediococcus pentosaceus as Well as Lactic Acid and Bacteriocin-like Inhibitory Substances (BLIS) Production

Pediococcus pentosaceus was cultivated in MRS medium supplemented or not with polydextrose under different conditions in order to evaluate its effect on cell growth, lactic acid and bacteriocin-like inhibitory substance (BLIS) production. Independent variables were pH (4.0, 5.0, 6.0), rotational speed (50, 100, 150 rpm), polydextrose concentration (0.5, 1.0, 1.5%) and temperature (25, 30, 35 °C), while cell concentration and productivity after 24 h, maximum specific growth rate, specific rate of substrate (glucose) consumption, volumetric and specific lactic acid productivities, yields of biomass and lactic acid on consumed substrate were the dependent . The maximum cell concentration (10.24 ± 0.16 gX L−1) and productivity (0.42 ± 0.01gX L−1 h−1) were achieved at pH 6.0, 35 °C, 150 rpm using 1.5% polydextrose, while the maximum specific growth rate (0.99 ± 0.01 h−1) and yield of biomass (2.96 ± 0.34gX gS−1) were achieved at the same pH and polydextrose concentration, but at 25 °C and 50 rpm. The specific substrate consumption rate (0.09 ± 0.02 gS gX−1 h−1) and the volumetric lactic acid productivity (0.44 ± 0.02 gP L−1 h−1) were maximized at pH 6.0, 35 °C, 50 rpm and 0.5% polydextrose. BLIS produced in this last run displayed the highest antibacterial activity against Escherichia coli, while the same activity was displayed against Enterococcus faecium using 1.5% polydextrose. These results appear to be quite promising in view of possible production of this BLIS as an antibacterial agent in the food industr

Cladosporium tenuissimum URM 7803: a promising new β-galactosidase producer

The Cladosporium genus, defined by Link in 1816, is one of the largest and most heterogeneous Hyphomycetes genus. It comprises more than 189 species still rarely explored biotechnologically. One of the most studied microbial enzymes, -galactosidase is a glycoside hydrolase enzyme that catalyzes the hydrolysis of -galactosides into monosaccharides through the breaking of a glycosidic bond. Recently, new studies comprising new microbial sources of -galactosidase, presenting biotechnologically interesting characteristics, have been encouraged. In this context, the present study evaluated the production of -galactosidase by a new isolate of Cladosporium tenuissimum. A C. tenuissimum inoculum was prepared adding 107 spore/mL in sterile saline solution 0.85% (w/v) NaCl containing 0.01% (w/v) Tween 80 and added to fermentation medium for enzyme production. The fermentation medium, composed of (% w/v): lactose (2), peptone (0.4), yeast extract (0.4) and salts (KH2PO4 (0.2), Na2HPO4.12H2O (0.8) and MgSO4.7H2O (0.025), pH 6.5, was maintained at 28° C and 180 rpm for 13 days. One sample (50 mL erlenmeyer) was removed every 24 hours and -galactosidase activity was evaluated using ONPG (ortho-Nitrophenyl--galactoside) method. The results showed maximum -galactosidase production by C. tenuissimum URM 7803 on thirteenth day, displayed enzymatic activity of 462.13 U/mL. The C. tenuissimum URM 7803 isolate proved to be a powerful new -galactosidase producer with potential application for food processing.info:eu-repo/semantics/publishedVersio

Biotechnological production of the antimicrobial peptide by Pediococcus pentosaceus ATCC43200 in growth supplemented with polydextrose.

Pediocinas são moléculas catiônicas, pequenas, organizadas em porção N-terminal hidrofílica- YGNGV e porção C-terminal anfifílica variável. A termoestabilidade e ação antimicrobiana agregam características importantes a essas bacteriocinas da classe IIa. Por meio da biopreservação alimentar, a atividade anti-listeria tornou-se uma particularidade dessas biomoléculas. Dentre as bactérias ácido láticas (BALs) encontram-se diferentes cepas bioprodutoras de pediocinas com importantes propriedades biotecnológicas. O gênero de Pediococcus spp.define-se por cocos rearranjados em tétrades, catalase negativa e anaeróbios facultativos. Pediococcus pentosaceus e Pediococcus acidilactici são as principais espécies empregadas na otimização da biosíntese de pediocinas. Alguns estudos desenvolvem-se com isolados bacterianos provenientes de múltiplos nichos biológicos e promovem sistemas heterogênios para conduzir os processos fermentativos. O atual estudo reune as informações significantes a respeito de cultivo, modo de ação, variedade de micro-organismos produtores do gênero Pediococcus spp. e aplicações biotecnológicas, ênfase nas alimentares para melhor compreensão do tema proposto.Pediocins are cationic molecules, small, arranged in a hydrophilic N-terminal portion - YGNGV - and an amphiphilic C-terminal variable portion. The thermal stability and antimicrobial effect add important features to these bacteriocins of class IIa. Through food biopreservation, anti-listeria activity became a peculiarity of these biomolecules. Among the lactic-acid bacteria (LAB), are different pediocin bioproducer strains with important biotechnological properties. Pediococcus spp. are characterized by coccus shaped bacteria arranged in tetrads, Gram-positive, non-motile, non-spore forming, catalase negative and facultative anaerobes. Pediococcus pentosaceus and Pediococcus acidilactici are the main species used in the optimization of the pediocins biosynthesis. Some studies have been developed with bacteria isolates from multiple biological niches and promote heterogeneous systems to conduct fermentation processes. This review joins the most significant information about cultivation, mode of action, variability of bacteriocin producer microorganisms from Pediococcus spp. and the biotechnological application of these bacteriocins, with emphasis on food products

First report of collagenase production by Trichosporon sp. strain isolated from pollen of Amazonian bee (Melipona seminigra seminigra)

Trichosporon yeasts are widely employed to produce lipids, lipases, and aspartic peptidases, but there are no previous studies on collagenase production. This work aimed to select the best collagenase producing Amazonian Trichosporon strains. Moreover, a 2(3)-full factorial design (FFD) and a 2(2)-central composite design combined with Response Surface Methodology were applied to optimize production and find the best conditions for hydrolysis of type I bovine collagen. Most of the studied strains had some collagenolytic activity, but the selected one achieved the highest value (44.02 U) and a biomass concentration of 2.31 g/L. The best collagenase production conditions were 160 rpm of agitation, pH 5.5 and a substrate concentration of 4.0 g/L. The former experimental design showed that substrate concentration was the only statistically significant factor on both biomass concentration and collagenase activity, while the latter showed simultaneous effects of substrate concentration and pH on collagenolytic activity, which peaked at pH 5.5-6.4 and substrate concentration of 3.0-3.4 g/L. An additional 2(3)-FFD was finally used to optimize the conditions collagen hydrolysis, and pH 6, 25 degrees C and a substrate concentration of 7.5 (g/L) ensured the highest hydrolysis degree. This study is the first that describes optimized conditions of collagenase production by Trichosporon strains

Collagenolytic enzymes produced by fungi: a systematic review

Specific proteases capable of degrading native triple helical or denatured collagen have been required for many years and have a large spectrum of applications. There are few complete reports that fully uncover production, characterization and purification of fungi collagenases. In this review, authors searched through four scientific on line data bases using the following keywords (collagenolytic OR collagenase) AND (fungi OR fungus OR fungal) AND (production OR synthesis OR synthesize) AND (characterization). Scientific criteria were adopted in this review to classify found articles by score (from 0 to 10). After exclusion criteria, 21 articles were selected. None obtained the maximum of 10 points defined by the methodology, which indicates a deficiency in studies dealing simultaneously with production, characterization and purification of collagenase by fungi. Among microorganisms studied the non-pathogenic fungi Penicillium aurantiogriseum and Rhizoctonia solani stood out in volumetric and specific collagenase activity. The only article found that made sequencing of a true collagenase showed 100% homology with several metalloproteinases fungi. A clear gap in literature about collagenase production by fungi was verified, which prevents further development in the area and increases the need for further studies, particularly full characterization of fungal collagenases with high specificity to collagen.This work was supported by Fundação de Amparo à Ciência e Tecnologia do Estado de Pernambuco (FACEPE), ConselhoNacional de Desenvolvimento Científico e Tecnológico (CNPq) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)