scholarly journals Isolation, purification and characterization of mannan degrading enzyme from a new isolate Acinetobacter baummanni

2021 ◽  
Vol 924 (1) ◽  
pp. 012078
Author(s):  
D Muzaki ◽  
E Zubaidah ◽  
S Santoso ◽  
A Sutrisno
Keyword(s):  
Enzyme Activity ◽  
Guar Gum ◽  
Relative Activity ◽  
Purification And Characterization ◽  
Rrna Sequence ◽  
16S Rrna Sequence ◽  
Ammonium Sulphate Precipitation ◽  
Optimum Ph ◽  
16S Rrna Sequence Analysis

Abstract A mannan-degrading microbe was isolated from rotting porang tubers (Amorphophallus muelleri Blume). Molecular identification using 16S-rRNA sequence analysis revealed that the isolate showed 99.67% similarity with Acinetobacter baummanni. A crude enzyme from ammonium sulphate precipitation was used for preliminary characterization. The characterization results showed that the enzyme activity is optimum at 45 °C, and stable at 35-50 °C, while the optimum pH is 7, and stable at pH 5-7. The substrate with the highest relative activity was found in guar gum which was 137.512%. The enzyme activity was inhibited by Ca, Na, K ions, and increased by Mn2+ ions.

scholarly journals Partial purification and characterization of an endo-α-N-acetylgalactosaminidase from the culture medium of Streptomyces sp. OH-11242

1992 ◽  
Vol 288 (2) ◽  
pp. 475-482 ◽  
Author(s):  
I Ishii-Karakasa ◽  
H Iwase ◽  
K Hotta ◽  
Y Tanaka ◽  
S Omura
Keyword(s):  
Enzyme Activity ◽  
Culture Medium ◽  
Gel Filtration ◽  
Partial Purification ◽  
Gastric Mucus ◽  
Purification And Characterization ◽  
Streptomyces Sp ◽  
Optimum Ph ◽  
New Type

For the purification of a new type of endo-alpha-N-acetylgalactosaminidase from the culture medium of Streptomyces sp. OH-11242 (endo-GalNAc-ase-S) [Iwase, Ishii, Ishihara, Tanaka, Omura & Hotta (1988) Biochem. Biophys. Res. Commun. 151, 422-428], a method for assaying enzyme activity was established. Using purified pig gastric mucus glycoprotein (PGM) as the substrate, oligosaccharides liberated from PGM were pyridylaminated, and the reducing terminal sugars of oligosaccharides larger than Gal beta 1-3GalNAc were analysed by h.p.1.c. The crude enzyme of endo-GalNAc-ase-S was prepared as an 80% (w/v) ammonium sulphate precipitate from the concentrated culture medium. The enzyme was partially purified by gel chromatofocusing and subsequent DEAE-Toyopearl chromatography. Endo-enzyme activity eluted around pI 4.8 on a gel chromatofocusing column and eluted with 0.19-0.25 M-NaCl on a DEAE-Toyopearl column. In the enzyme fraction obtained, no exo-glycosidases or proteases could be detected. The molecular mass of the enzyme was estimated as 105 kDa by gel filtration, and the optimum pH was 5.5. Endo-GalNAc-ase-S hydrolysed the O-glycosidic linkage between GalNAc and Ser (Thr) in 3H-labelled and unlabelled asialofetuin, liberating both the disaccharide (Gal beta 1-3GalNAc) and the tetrasaccharide [Gal beta 1-3 (Gal beta 1-4GlcNAc beta 1-6)GalNAc]. When endo-alpha-N-acetylgalactosaminidase from Alcaligenes sp. (endo-GalNac-ase-A) was incubated with 3H-labelled and unlabelled asialofetuin, only the disaccharide (Gal beta 1-3GalNAc) was liberated.

scholarly journals Optimization of Growth Conditions for Purification and Production of L-Asparaginase bySpirulina maxima

2016 ◽  
Vol 2016 ◽  
pp. 1-7 ◽  
Author(s):  
Hanaa H. Abd El Baky ◽  
Gamal S. El Baroty
Keyword(s):  
Enzyme Activity ◽  
Gel Filtration ◽  
Growth Conditions ◽  
Partial Purification ◽  
Specific Enzyme ◽  
Purification And Characterization ◽  
Spirulina Maxima ◽  
Optimum Ph ◽  
Biomass Yields

L-asparaginase (L-AsnA) is widely distributed among microorganisms and has important applications in medicine and in food technology sectors. Therefore, the ability of the production, purification, and characterization of AsnA fromSpirulina maxima(SM) were tested. SM cultures grown in Zarrouk medium containing different N2(in NaNO3form) concentrations (1.25, 2.50, and 5.0 g/L) for 18 days contained a significant various quantity of dry biomass yields and AsnA enzyme levels. MS L-AsnA activity was found to be directly proportional to the N2concentration. The cultures of SM at large scales (300 L medium, 5 g/L N2) showed a high AsnA enzyme activity (898 IU), total protein (405 mg/g), specific enzyme activity (2.21 IU/mg protein), and enzyme yield (51.28 IU/L) compared with those in low N2cultures. The partial purification of crude MS AsnA enzyme achieved by 80% ammonium sulfate AS precipitated and CM-Sephadex C-200 gel filtration led to increases in the purification of enzyme with 5.28 and 10.91 times as great as that in SM crude enzymes. Optimum pH and temperature of purified AsnA for the hydrolyzate were 8.5 and 37 ± 0.2°C, respectively. To the best of our knowledge, this is the first report on L-asparaginase production inS. maxima.

scholarly journals Characterization of Yellow Pigmented Bacteria Associated with Gracilaria sp.

2020 ◽  
Vol 2 (2) ◽  
Author(s):  
Arina Tri Lunggani ◽  
Susianna Purwantisari ◽  
Siti Nur Jannah
Keyword(s):  
Room Temperature ◽  
Research Result ◽  
Growth Media ◽  
Bacterial Isolation ◽  
Rrna Sequence ◽  
16S Rrna Sequence ◽  
Kinship Analysis ◽  
16S Rrna Sequence Analysis ◽  
Pigmented Bacteria

Research on the kinship analysis of endophytic bacterial  isolated from Gracillaria sp has been carried out. The presence of bacteria associated with Gracilaria sp. has enabled the use of these bacteria as a source of new bioactive compounds, such as biopigments. The research aims to isolated bacteria from Gracilaria sp., screened their symbiont bacteria that could potentially produce pigments. Sampling Gracilaria sp. conducted in the waters of the Island of  Karimunjawa, Jepara. Furthermore, bacterial isolation was carried out, screening for pigment-producing bacteria and 16S rRNA sequence analysis. Research result showed that the symbiont bacteria isolate TK 373 produced consistent pigments after several regenerations, in several types of growth media incubated at room temperature. The results of 16S rDNA identification showed that the TK 373 isolate had the closest relationship with  Pseudoalteromonas sp. with  98.72 % homology.

scholarly journals Characterization of Bacterial Community of Rumen in Dairy Cows With Laminitis

2021 ◽  
Author(s):  
Jian Gao ◽  
Bo Liu ◽  
Shuang Li ◽  
Ruiying Mu ◽  
Naisheng Zhang ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Rumen Fluid ◽  
Economic Loss ◽  
Control Group ◽  
Rrna Sequence ◽  
16S Rrna Sequence ◽  
16S Rrna Sequence Analysis ◽  
Clostridium Papyrosolvens ◽  
Ruminal Microbiota

Abstract Background: Laminitis- an inflammation of lamella, could cause great economic loss to dairy industry, which has attracted wide attention around the world. In recent years, microbiota is considered as one of the vital parts that played significant role in various diseases processes. However, current studies are far from sufficient. Aim of this study is to explore the characteristics of ruminal microbiota in laminitis cows. Methods: The serum of bovines with or without laminitis was collected to detect concentrations of lipopolysaccharide (LPS), lactic acid, and histamine, and the ruminal fluid was collected for 16S rRNA sequence analysis. Results: The results showed that a significant increase in LPS and lactic acid levels in laminitis group comparing to control group cows. In addition, the higher abundance of bacteria that -riches acid-enhancing metabolites, namely, Candidatus Saccharimonas, Saccharofermentans, Erysipelotrichaceae UCG-009, Erysipelotrichaceae UCG-008, Clostridium papyrosolvens and Ruminococcaceae bacterium AE2021 were detected in the rumen fluid from laminitis bovines. Conclusions: This article confirmed that difference of rumen microbiota were occurred in rumen between health and laminitis bovines. The elevated abundance of bacteria that riches acid- enhancing metabolites, as well as increased the concentration of lactic acid and LPS could be harmful factors to bovines and increase risks of laminitis.

Purification and characterization of a new α-amylase of intermediate thermal stability from the yeast Lipomyces kononenkoae

1995 ◽  
Vol 73 (1-2) ◽  
pp. 41-49 ◽  
Author(s):  
José Antonio Prieto ◽  
Bernardo Roque Bort ◽  
Javier Martínez ◽  
Francisca Randez-Gil ◽  
Pascual Sanz ◽  
...  
Keyword(s):  
Thermal Stability ◽  
Enzyme Activity ◽  
Ammonium Sulphate ◽  
Soluble Starch ◽  
Affinity Binding ◽  
Purification And Characterization ◽  
Lipomyces Kononenkoae ◽  
Optimum Ph

A new α-amylase from the extracellular culture of the yeast Lipomyces kononenkoae CBS 5608 has been purified to homogeneity by ammonium sulphate treatment, affinity binding on cross-linked starch, and DEAE–Biogel A chromatography. The enzyme was monomeric, with an apparent Mr of 76 kilodaltons, pI < 3.5, and optimum pH 4.5–5.0, and exhibited intermediate thermal stability. The temperature for optimal enzyme activity was 70 °C. It is a glycoprotein with both N- and O-linked sugars. Kinetic analyses indicate that the enzyme has an endoamylolytic mechanism. The kM for soluble starch was 0.80 g∙L−1 and the kcat was 622∙s−1. Keywords: α-amylase, glycosylation, intermediate thermal stability, Lipomyces kononenkoae.

scholarly journals Pseudomonas eucalypticola sp. nov., a producer of antifungal agents isolated from Eucalyptus dunnii leaves

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yujing Liu ◽  
Zhang Song ◽  
Hualong Zeng ◽  
Meng Lu ◽  
Weiyao Zhu ◽  
...  
Keyword(s):  
Dna Hybridization ◽  
Antifungal Agents ◽  
Novel Species ◽  
Phytopathogenic Fungi ◽  
Strictly Aerobic ◽  
Rrna Sequence ◽  
16S Rrna Sequence ◽  
Eucalyptus Dunnii ◽  
16S Rrna Sequence Analysis

AbstractPseudomonas are ubiquitously occurring microorganisms and are known for their ability to produce antimicrobials. An endophytic bacterial strain NP-1 T, isolated from Eucalyptus dunnii leaves, exhibits antifungal properties against five tested phytopathogenic fungi. The strain is a Gram-negative rod-shaped bacterium containing a single polar flagellum. It is strictly aerobic, grows at 4–37 °C, 2–5% NaCl, and pH 3–7. The 16S rRNA sequence analysis showed that NP-1 T belongs to the Pseudomonas genus. Phylogenetic analysis based on four concatenated partial genes (16S rDNA, gyrB, rpoB and rpoD) and the phylogenomic tree indicated that NP-1 T belongs to Pseudomonas fluorescens lineage but is distinct from any known Pseudomonas species. The G + C mol % of NP-1 T genome is 63.96, and the differences between NP-1 T and related species are larger than 1. The digital DNA-DNA hybridization and tetranucleotide signatures are 23.8 and 0.97, which clearly separates strain NP-1 T from its closest neighbours, Pseudomonas coleopterorum and Pseudomonas rhizosphaerae. Its phenotypic and chemotaxonomic features confirmed its differentiation from related taxa. The results from this polyphasic approach support the classification of NP-1 T as a novel species of Pseudomonas, and the name of Pseudomonas eucalypticola is thus proposed for this strain, whose type is NP-1 T (= CCTCC M2018494T = JCM 33572 T).

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