scholarly journals Biochemical Properties of Crude Extracellular Proteases from Chromohalobacter salexigens BKL5 and Micrococcus luteus 11A

2017 ◽  
Vol 21 (1) ◽  
pp. 56
Author(s):  
Ayu Ashari Achmad ◽  
M. Saifur Rohman ◽  
Irfan D. Prijambada
Keyword(s):  
Enzymatic Activity ◽  
Specific Activity ◽  
Micrococcus Luteus ◽  
Skim Milk ◽  
Biochemical Properties ◽  
Extracellular Protease ◽  
Bacterial Strains ◽  
Extracellular Proteases ◽  
Metal Cofactor ◽  
Chromohalobacter Salexigens

In this work, we have reported an enzymatic activity and biochemical properties of extracellular proteases from Chromohalobacter salexigens BKL5 and Micrococcus luteus 11A. C. salexigens BKL5 and M. luteus 11A were previously isolated from Bledug Kuwu mud volcano and dietary industry wastewater treatment, respectively. Both bacterial strains were able to produce extracellular proteases, when grown on minimal agar medium supplemented with 1% of skim milk. Proteolytic indexes of C. salexigens BKL5 and M. luteus 11A were 2.5±0.14 and 2.9±0.42, respectively. Both extracellular proteases exhibited optimum enzymatic activity at pH 7, with specific activity of C. salexigens BKL5 was 13.3% higher than that of M. luteus 11A. Optimum temperature for enzymatic activity of both proteases was 45°C. Metal cofactor preferences assay showed that extracellular protease from C. salexigens BKL5 preferred Zn2+, meanwhile extracellular protease from M. luteus 11A mainly preferred Ca2+ ion. Metal cofactor preferences assay also suggested that crude extracellular protease from C. salexigens BKL5 was categorized as metalloprotease, meanwhile crude extracellular protease of M. luteus 11A was common neutral protease. The enzymatic stability assay against various salt concentrations showed that crude extracellular protease from C. salexigens BKL5 was more stable than that of M. luteus 11A.

scholarly journals ENHANCED ENZYMATIC ACTIVITY OF STREPTOMYCES GRISEOPLANUS L-ASPARGINASE VIA ITS INCORPORATION IN AN OIL-BASED NANOCARRIER

2020 ◽  
pp. 203-210
Author(s):  
ABEER A. EL-HADI ◽  
HANAN MOSTAFA AHMED ◽  
RANIA A. ZAKI ◽  
AMIRA MOHAMED MOHSEN
Keyword(s):  
Thermal Stability ◽  
Oleic Acid ◽  
Enzymatic Activity ◽  
Specific Activity ◽  
Residual Activity ◽  
Tween 80 ◽  
Biochemical Properties ◽  
Lymphocytic Leukemia ◽  
Tween 20 ◽  
Ph Range

Objective: L-asparaginase (L-asp) is a vital enzyme used as a therapeutic agent in combination with other drugs in the treatment of acute lymphoma, melanosarcoma and lymphocytic leukemia. Immobilization of enzymes through loading on nanoemulsion (NE) results in some advantages such as enhancing their stability and increasing their resistance to proteases. Aim of the present study is to formulate L-asp loaded nanoemulsion to enhance its efficiency and thermal stability. Methods: Nanoemulsion loaded with L-asp crude extract (specific activity 13.23U/mg protein) was prepared employing oleic acid as oil, tween 20/tween 80 as surfactants and propylene glycol (PG) as co-surfactant. L-asp loaded NE underwent several thermodynamic stability studies and the optimized formulae were further examined for their biochemical properties and thermal stability. Results The developed formulations were spherical in shape and their sizes were in the nanometric dimensions with negatively charged zeta potential values. Upon comparing the enzyme activity of L-asp loaded NE employing tween 20 (F1) or tween80 (F4) at different concentrations, the results revealed that F4 NE showed higher enzymatic activity [323 U/ml] compared to F1 NE [197 U/ml] at the same concentration. The nanosized immobilized L-asp was more stable in the pH range from 8 to 8.5 as compared to free L-asp. The immobilized enzyme preserved about 59.11% of its residual activity at 50 °C; while free L-asp preserved about 33.84%. Conclusion: In the view of these results, NE composed of oleic acid, tween 80 and PG represents a promising dosage form for enhancing the activity and stability of Streptomyces griseoplanus L-asp.

scholarly journals Substrate preferences, phylogenetic and biochemical properties of proteolytic bacteria present in the digestive tract of Nile tilapia

2021 ◽  
Author(s):  
Tanim Jabid Hossain ◽  
Mukta Das ◽  
Ferdausi Ali ◽  
Sumaiya Islam Chowdhury ◽  
Subrina Akter Zedny
Keyword(s):  
Nile Tilapia ◽  
Molecular Taxonomy ◽  
Skim Milk ◽  
Biochemical Properties ◽  
Protease Production ◽  
Rrna Gene ◽  
Bacterial Strains ◽  
Substrate Preferences ◽  
Proteolytic Bacteria ◽  
Excellent Source

Abstract Vertebrate intestine appears an excellent source of proteolytic bacteria for industrial and probiotic use. We therefore aimed obtaining the gut-associated proteolytic species of Nile tilapia. We’ve isolated twenty six bacterial strains from its intestinal tract, seven of which showed exoprotease activity with the formation of clear halos on skim milk. Their depolymerization ability was further assessed on three distinct proteins including casein, gelatin and albumin. All the isolates could successfully hydrolyze the three substrates indicating relatively broad specificity of their secreted proteases. Molecular taxonomy and phylogeny of the proteolytic isolates were determined based on their 16S rRNA gene barcoding which suggested that the seven strains belong to three phyla viz. Firmicutes, Proteobacteria and Actinobacteria, distributed across the genera Priestia, Citrobacter, Pseudomonas, Stenotrophomonas, Burkholderia, Providencia and Micrococcus. The isolates were further characterized by a comprehensive study of their morphological, cultural, cellular and biochemical properties which were consistent with the phylogenetic annotations. To reveal their proteolytic capacity alongside substrate preferences, enzyme-production was determined by the diffusion assay. The Pseudomonas, Stenotrophomonas and Micrococcus isolates appeared most promising with maximum protease production on casein, gelatin and albumin media respectively. Our findings present valuable insights into the phylogenetic and biochemical properties of gut-associated proteolytic strains of Nile tilapia.

scholarly journals Substrate preferences, phylogenetic and biochemical properties of proteolytic bacteria present in the digestive tract of Nile tilapia (Oreochromis niloticus)

2021 ◽  
Vol 7 (4) ◽  
pp. 528-545
Author(s):  
Tanim Jabid Hossain ◽  
◽  
Mukta Das ◽  
Ferdausi Ali ◽  
Sumaiya Islam Chowdhury ◽  
...  
Keyword(s):  
Oreochromis Niloticus ◽  
Nile Tilapia ◽  
Molecular Taxonomy ◽  
Skim Milk ◽  
Biochemical Properties ◽  
Protease Production ◽  
Rrna Gene ◽  
Bacterial Strains ◽  
Substrate Preferences ◽  
Proteolytic Bacteria

<abstract> <p>Vertebrate intestine appears to be an excellent source of proteolytic bacteria for industrial and probiotic use. We therefore aimed at obtaining the gut-associated proteolytic species of Nile tilapia (<italic>Oreochromis niloticus</italic>). We have isolated twenty six bacterial strains from its intestinal tract, seven of which showed exoprotease activity with the formation of clear halos on skim milk. Their depolymerization ability was further assessed on three distinct proteins including casein, gelatin, and albumin. All the isolates could successfully hydrolyze the three substrates indicating relatively broad specificity of their secreted proteases. Molecular taxonomy and phylogeny of the proteolytic isolates were determined based on their 16S rRNA gene barcoding, which suggested that the seven strains belong to three phyla viz. Firmicutes, Proteobacteria, and Actinobacteria, distributed across the genera <italic>Priestia</italic>, <italic>Citrobacter</italic>, <italic>Pseudomonas</italic>, <italic>Stenotrophomonas</italic>, <italic>Burkholderia</italic>, <italic>Providencia</italic>, and <italic>Micrococcus</italic>. The isolates were further characterized by a comprehensive study of their morphological, cultural, cellular and biochemical properties which were consistent with the phylogenetic annotations. To reveal their proteolytic capacity alongside substrate preferences, enzyme-production was determined by the diffusion assay. The <italic>Pseudomonas</italic>, <italic>Stenotrophomonas</italic> and <italic>Micrococcus</italic> isolates appeared to be most promising with maximum protease production on casein, gelatin, and albumin media respectively. Our findings present valuable insights into the phylogenetic and biochemical properties of gut-associated proteolytic strains of Nile tilapia.</p> </abstract>

scholarly journals Substrate preferences, phylogenetic and biochemical properties of proteolytic bacteria present in the digestive tract of Nile tilapia

2021 ◽  
Author(s):  
Tanim Jabid Hossain ◽  
Mukta Das ◽  
Ferdausi Ali ◽  
Sumaiya Islam Chowdhury ◽  
Subrina Akter Zedny
Keyword(s):  
Nile Tilapia ◽  
Molecular Taxonomy ◽  
Skim Milk ◽  
Biochemical Properties ◽  
Protease Production ◽  
Rrna Gene ◽  
Bacterial Strains ◽  
Substrate Preferences ◽  
Proteolytic Bacteria ◽  
Excellent Source

Vertebrate intestine appears an excellent source of proteolytic bacteria for industrial and probiotic use. We therefore aimed obtaining the gut-associated proteolytic species of Nile tilapia. We have isolated twenty six bacterial strains from its intestinal tract, seven of which showed exoprotease activity with the formation of clear halos on skim milk. Their depolymerization ability was further assessed on three distinct proteins including casein, gelatin and albumin. All the isolates could successfully hydrolyze the three substrates indicating relatively broad specificity of their secreted proteases. Molecular taxonomy and phylogeny of the proteolytic isolates were determined based on their 16S rRNA gene barcoding which suggested that the seven strains belong to three phyla viz. Firmicutes, Proteobacteria and Actinobacteria, distributed across the genera Priestia, Citrobacter, Pseudomonas, Stenotrophomonas, Burkholderia, Providencia and Micrococcus. The isolates were further characterized by a comprehensive study of their morphological, cultural, cellular and biochemical properties which were consistent with the phylogenetic annotations. To reveal their proteolytic capacity alongside substrate preferences, enzyme-production was determined by the diffusion assay. The Pseudomonas, Stenotrophomonas and Micrococcus isolates appeared most promising with maximum protease production on casein, gelatin and albumin media respectively. Our findings present valuable insights into the phylogenetic and biochemical properties of gut-associated proteolytic strains of Nile tilapia.

Cloning, expression, and enzymatic activity analysis of strawberry S-adenosyl-L-metnionine synthetase gene FaSAMS1

2016 ◽  
Vol 6 (1) ◽  
pp. 841-845
Author(s):  
Shu-Fang Wang ◽  
Li-Na Hua ◽  
Xiaofeng Liu ◽  
Yuanyue Shen
Keyword(s):  
Enzymatic Activity ◽  
Specific Activity ◽  
Reaction System ◽  
Biochemical Properties ◽  
Activity Analysis ◽  
Yeast Expression ◽  
Strawberry Fruit ◽  
Adenosyl Methionine ◽  
Strawberry Fruit Ripening ◽  
Expression Strain

The S-adenosyl methionine synthetase gene FaSAMS1 is involved in the regulation of strawberry fruit ripening, however, the biochemical properties of FaSAMS1 protein remain unclear. Here, a coding cDNA sequence of FaSAMS1 was cloned by RT-PCR and inserted into a recombinant yeast expression vector pPICZA, then transformed into the yeast expression strain X-33. The fusion protein FaSAMS1 was induced, expressed, and purified. The enzymatic activity analysis of FaSAMS1 showed that the reaction system contains 0.09 mg of FaSAMS1, protein concentration reached at 0.454 mg·mL-1, the activity of the enzyme was 0.32 U, specific activity was 3.56 U·mg-1. This study has provided biochemical evidence for the involvement of ethylene in the regulation of strawberry fruit ripening. 

scholarly journals Partial purification, characterization and immobilization of a novel lipase from a native isolate of Lactobacillus fermentum

2021 ◽  
Author(s):  
Foruzan Fathi ◽  
Rouha Kasra-Kermanshahi ◽  
Zahra Moosavi-Nejad ◽  
Elahe Mobarak Qamsari
Keyword(s):  
Enzymatic Activity ◽  
Ammonium Sulfate ◽  
High Efficiency ◽  
Specific Activity ◽  
Lipolytic Activity ◽  
Physical Adsorption ◽  
Lactobacillus Fermentum ◽  
Partial Purification ◽  
Bacterial Strains ◽  
Chitosan Beads

Background and Objectives: Due to the widespread use of lipase enzymes in various industries, finding native lipase pro- ducing microorganisms is of great value and importance. In this study, screening of lipase-producing lactobacilli from native dairy products was performed. Materials and Methods: Qualitative evaluation of lipolytic activity of lipase-producing lactobacilli was performed in differ- ent media containing olive oil. A clear zone observation around the colonies indicated the lipolytic activity. The strain with the highest enzymatic activity was identified. Determination of optimal pH and temperature of lipase activity was measured by spectrophotometry using p-nitrophenyl acetate (ρ-NPA) substrate. Partial purification of lipase enzyme was performed using 20-90% saturation ammonium sulfate. Eventually, lipase was immobilized by physical adsorption on chitosan beads. Results: Among screened lipolytic bacterial strains, one sample (5c isolate) which showed the highest enzymatic activity (5329.18 U/ml) was close to Lactobacillus fermentum. During characterization, the enzyme showed maximum activity in Tris-HCl buffer with pH 7, while remaining active over a temperature range of 5°C to 40°C. The results of the quantitative assay demonstrated that the fraction precipitated in ammonium sulfate at 20% saturation has the highest amount of lipolytic activity, with a specific activity of 22.0425 ± 3.6 U/mg. Purification folds and yields were calculated as 8.73 and 44%, respec- tively. Eventually, the enzyme was immobilized by physical adsorption on chitosan beads with a yield of 56.21%. Conclusion: The high efficiency of enzyme immobilization on chitosan beads indicates the suitability of this method for long-term storage of new lipase from native 5c isolate.

THE EFFECT OF BACTERIAL STRAINS ON THE ENZYMATIC ACTIVITY OF SOILS IN IMPACT ZONE OF ATAMAN LAKE

2020 ◽  
Author(s):  
V.V. Zinchenko ◽  
◽  
E.S Fedorenko ◽  
A.V Gorovtsov ◽  
T.M Minkina ◽  
...  
Keyword(s):  
Enzymatic Activity ◽  
Dehydrogenase Activity ◽  
Contaminated Soil ◽  
Model Experiment ◽  
Impact Zone ◽  
Bacterial Strains ◽  
The Impact

As a result of the model experiment, an increase in the enzymatic activity of meadow chernozem of the impact zone of Ataman Lake with the introduction of a strains mixture of metal-resistant microorganisms into the soil was established. The experiment has shown that the application of bacterial strains increases the dehydrogenase activity of contaminated soil by 51.8% compared to the variant without remediation

scholarly journals Biochemical characterization of specific Alanine Decarboxylase (AlaDC) and its ancestral enzyme Serine Decarboxylase (SDC) in tea plants (Camellia sinensis)

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Peixian Bai ◽  
Liyuan Wang ◽  
Kang Wei ◽  
Li Ruan ◽  
Liyun Wu ◽  
...  
Keyword(s):  
Gene Duplication ◽  
Camellia Sinensis ◽  
Biochemical Characterization ◽  
Specific Activity ◽  
Protein Sequences ◽  
Biochemical Properties ◽  
High Similarity ◽  
New Gene ◽  
Tea Plants

Abstract Background Alanine decarboxylase (AlaDC), specifically present in tea plants, is crucial for theanine biosynthesis. Serine decarboxylase (SDC), found in many plants, is a protein most closely related to AlaDC. To investigate whether the new gene AlaDC originate from gene SDC and to determine the biochemical properties of the two proteins from Camellia sinensis, the sequences of CsAlaDC and CsSDC were analyzed and the two proteins were over-expressed, purified, and characterized. Results The results showed that exon-intron structures of AlaDC and SDC were quite similar and the protein sequences, encoded by the two genes, shared a high similarity of 85.1%, revealing that new gene AlaDC originated from SDC by gene duplication. CsAlaDC and CsSDC catalyzed the decarboxylation of alanine and serine, respectively. CsAlaDC and CsSDC exhibited the optimal activities at 45 °C (pH 8.0) and 40 °C (pH 7.0), respectively. CsAlaDC was stable under 30 °C (pH 7.0) and CsSDC was stable under 40 °C (pH 6.0–8.0). The activities of the two enzymes were greatly enhanced by the presence of pyridoxal-5′-phosphate. The specific activity of CsSDC (30,488 IU/mg) was 8.8-fold higher than that of CsAlaDC (3467 IU/mg). Conclusions Comparing to CsAlaDC, its ancestral enzyme CsSDC exhibited a higher specific activity and a better thermal and pH stability, indicating that CsSDC acquired the optimized function after a longer evolutionary period. The biochemical properties of CsAlaDC might offer reference for theanine industrial production.

scholarly journals Expression of novel acidic lipase from Micrococcus luteus in Pichia pastoris and its application in transesterification

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Selfela Restu Adina ◽  
Antonius Suwanto ◽  
Anja Meryandini ◽  
Esti Puspitasari
Keyword(s):  
Fatty Acid ◽  
Pichia Pastoris ◽  
Specific Activity ◽  
Signal Sequence ◽  
Micrococcus Luteus ◽  
Acid Methyl Ester ◽  
Industrial Applications ◽  
Expression Level ◽  
Lipase Gene ◽  
Acidic Lipase

Abstract Background Lipases are promising biocatalysts for industrial applications and attract attention to be explored. A novel acidic lipase has been isolated from the lipolytic bacteria Micrococcus luteus EMP48-D (LipEMP48-D) screened from tempeh. The lipase gene had previously been overexpressed in Escherichia coli BL21, but the expression level obtained was relatively low. Here, to improve the expression level, the lipase gene was cloned to Pichia pastoris. We eliminated the native signal sequence of M. luteus and replaced it with α-mating factor (α-MF) signal sequence. We also optimized and synthesized the lipase gene based on codon preference in P. pastoris. Results LipEMP48-D lipase was expressed as an extracellular protein. Codon optimization has been conducted for 20 codons, with the codon adaption index reaching 0.995. The highest extracellular lipase activity obtained reached 145.4 ± 4.8 U/mg under AOX1 promoter in P. pastoris KM71 strain, which was 9.7-fold higher than the previous activity in E. coli. LipEMP48-D showed the highest specific activity at pH 5.0 and stable within the pH range 3.0–5.0 at 40 °C. LipEMP48-D also has the capability of hydrolyzing various long-chain triglycerides, particularly olive oil (100%) followed by sunflower oil (88.5%). LipEMP48-D exhibited high tolerance for various polar organic solvents with low log P, such as isopropanol (115.7%) and butanol (114.6%). The metal ions (Na+, K+, Ca2+, Mg2+, Mn+) decreased enzyme activity up to 43.1%, while Fe2+ increased relative activity of enzymes up to 200%. The conversion of free fatty acid (FFA) into fatty acid methyl ester (FAME) was low around 2.95%. Conclusions This study was the first to report overexpression of Micrococcus lipase in yeast. The extracellular expression of this acidic lipase could be potential for biocatalyst in industrial fields, especially organic synthesis, food industry, and production of biodiesel.

scholarly journals Smells Like Teen Spirit—A Model to Generate Laundry-Associated Malodour In Vitro

2021 ◽  
Vol 9 (5) ◽  
pp. 974
Author(s):  
Marc-Kevin Zinn ◽  
Marco Singer ◽  
Dirk Bockmühl
Keyword(s):  
Volatile Compounds ◽  
Micrococcus Luteus ◽  
Dimethyl Disulfide ◽  
Bacterial Strains ◽  
Dimethyl Trisulfide ◽  
Model Based ◽  
Washing Machines ◽  
Staphylococcus Hominis

Although malodour formation on textiles and in washing machines has been reported to be a very relevant problem in domestic laundry, the processes leading to bad odours have not been studied intensively. In particular, the smell often described as “wet-and-dirty-dustcloth-like malodour” had not been reproduced previously. We developed a lab model based on a bacterial mixture of Micrococcus luteus, Staphylococcus hominis, and Corynebacterium jeikeium, which can produce this odour type and which might allow the detailed investigation of this problem and the development of counteractions. The model uses bacterial strains that have been isolated from malodourous textiles. We could also show that the three volatile compounds dimethyl disulfide, dimethyl trisulfide, and indole contribute considerably to the “wet-fabric-like” malodour. These substances were not only found to be formed in the malodour model but have already been identified in the literature as relevant malodourous substances.

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