scholarly journals Isolation and Characterization of Mutants of the Methylotrophic Yeast,Candida boidiniiS2 That Are Impaired in Growth on Peroxisome-Inducing Carbon Sources

1995 ◽  
Vol 59 (5) ◽  
pp. 869-875 ◽  
Author(s):  
Yasuyoshi Sakai ◽  
Hideaki Matsuo ◽  
Kai-Ze He ◽  
Atsushi Saiganji ◽  
Hiroya Yurimoto ◽  
...  
Keyword(s):  
Carbon Sources ◽  
Methylotrophic Yeast ◽  
Isolation And Characterization

scholarly journals Isolation and Characterization of a Genetically Tractable Photoautotrophic Fe(II)-Oxidizing Bacterium, Rhodopseudomonas palustris Strain TIE-1

2005 ◽  
Vol 71 (8) ◽  
pp. 4487-4496 ◽  
Author(s):  
Yongqin Jiao ◽  
Andreas Kappler ◽  
Laura R. Croal ◽  
Dianne K. Newman
Keyword(s):  
Bacterial Species ◽  
Rhodopseudomonas Palustris ◽  
Carbon Sources ◽  
Integral Membrane Protein ◽  
Transposition Frequency ◽  
Isolation And Characterization ◽  
Abc Transport ◽  
A Cell ◽  
Random Insertion

ABSTRACT We report the isolation and characterization of a phototrophic ferrous iron [Fe(II)]-oxidizing bacterium named TIE-1 that differs from other Fe(II)-oxidizing phototrophs in that it is genetically tractable. Under anaerobic conditions, TIE-1 grows photoautotrophically with Fe(II), H2, or thiosulfate as the electron donor and photoheterotrophically with a variety of organic carbon sources. TIE-1 also grows chemoheterotrophically in the dark. This isolate appears to be a new strain of the purple nonsulfur bacterial species Rhodopseudomonas palustris, based on physiological and phylogenetic analysis. Fe(II) oxidation is optimal at pH 6.5 to 6.9. The mineral products of Fe(II) oxidation are pH dependent: below pH 7.0 goethite (α-FeOOH) forms, and above pH 7.2 magnetite (Fe3O4) forms. TIE-1 forms colonies on agar plates and is sensitive to a variety of antibiotics. A hyperactive mariner transposon is capable of random insertion into the chromosome with a transposition frequency of ∼10−5. To identify components involved in phototrophic Fe(II) oxidation, mutants of TIE-1 were generated by transposon mutagenesis and screened for defects in Fe(II) oxidation in a cell suspension assay. Among approximately 12,000 mutants screened, 6 were identified that are specifically impaired in Fe(II) oxidation. Five of these mutants have independent disruptions in a gene that is predicted to encode an integral membrane protein that appears to be part of an ABC transport system; the sixth mutant has an insertion in a gene that is a homolog of CobS, an enzyme involved in cobalamin (vitamin B12) biosynthesis.

scholarly journals Isolation and characterization of omnipotent suppressors in the yeast Saccharomyces cerevisiae.

Genetics ◽  
1990 ◽  
Vol 124 (3) ◽  
pp. 515-522
Author(s):  
L P Wakem ◽  
F Sherman
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Genetic Analysis ◽  
Low Temperatures ◽  
Carbon Sources ◽  
Yeast Saccharomyces Cerevisiae ◽  
Hypertonic Medium ◽  
Isolation And Characterization ◽  
Wide Range ◽  
Chromosome Ii

Abstract Approximately 290 omnipotent suppressors, which enhance translational misreading, were isolated in strains of the yeast Saccharomyces cerevisiae containing the psi+ extrachromosomal determinant. The suppressors could be assigned to 8 classes by their pattern of suppression of five nutritional markers. The suppressors were further distinguished by differences in growth on paromomycin medium, hypertonic medium, low temperatures (10 degrees), nonfermentable carbon sources, alpha-aminoadipic acid medium, and by their dominance and recessiveness. Genetic analysis of 12 representative suppressors resulted in the assignment of these suppressors to 6 different loci, including the three previously described loci SUP35 (chromosome IV), SUP45 (chromosome II) and SUP46 (chromosome II), as well as three new loci SUP42 (chromosome IV), SUP43 (chromosome XV) and SUP44 (chromosome VII). Suppressors belonging to the same locus had a wide range of different phenotypes. Differences between alleles of the same locus and similarities between alleles of different loci suggest that the omnipotent suppressors encode proteins that effect different functions and that altered forms of each of the proteins can effect the same function.

scholarly journals Isolation and characterization of cold sensitive pex6 mutant of the methylotrophic yeast Hansenula polymorpha

2002 ◽  
Vol 18 (2) ◽  
pp. 131-134 ◽  
Author(s):  
V. Y. Nazarko ◽  
O. D. Pochapinsky ◽  
T. Y. Nazarko ◽  
O. V. Stasyk ◽  
A. A. Sibirny
Keyword(s):  
Hansenula Polymorpha ◽  
Methylotrophic Yeast ◽  
Isolation And Characterization ◽  
Cold Sensitive

Pichia pastoris as a host system for transformations

1985 ◽  
Vol 5 (12) ◽  
pp. 3376-3385
Author(s):  
J M Cregg ◽  
K J Barringer ◽  
A Y Hessler ◽  
K R Madden
Keyword(s):  
Pichia Pastoris ◽  
Auxotrophic Mutant ◽  
Methylotrophic Yeast ◽  
Fusion Procedure ◽  
Methylotrophic Yeast Pichia Pastoris ◽  
Isolation And Characterization ◽  
Mutant Host ◽  
Histidinol Dehydrogenase ◽  
His4 Gene

We developed a methylotrophic yeast, Pichia pastoris, as a host for DNA transformations. The system is based on an auxotrophic mutant host of P. pastoris which is defective in histidinol dehydrogenase. As a selectable marker, we isolated and characterized the P. pastoris HIS4 gene. Plasmid vectors which contained either the P. pastoris or the Saccharomyces cerevisiae HIS4 gene transformed the P. pastoris mutant host. DNA transfer was accomplished by a modified version of the spheroplast generation (CaCl2-polyethylene glycol)-fusion procedure developed for S. cerevisiae. In addition, we report the isolation and characterization of P. pastoris DNA fragments with autonomous replication sequence activity. Two fragments, PARS1 and PARS2, when present on plasmids increased transformation frequencies to 10(5)/micrograms and maintained the plasmids as autonomous elements in P. pastoris cells.

scholarly journals Isolation and characterization of soil actinobacteria as cellulolytic enzyme producer from Aceh Besar, Indonesia

2021 ◽  
Vol 22 (11) ◽  
Author(s):  
Lenni Fitri ◽  
MOHAMMAD ADZANNIE BESSANIA ◽  
NADIA SEPTI ◽  
SUHARTONO SUHARTONO
Keyword(s):  
Carbon Source ◽  
Carbon Sources ◽  
Morphological Characterization ◽  
Dilution Method ◽  
Cellulolytic Enzyme ◽  
Genus Streptomyces ◽  
Physiological Characterization ◽  
Isolation And Characterization ◽  
Index Value

Abstract. Fitri L, Bessania MA, Septi N, Suhartono S. 2021. Isolation and characterization of soil actinobacteria as cellulolytic enzyme producer from Aceh Besar, Indonesia. Biodiversitas 22: 5169-5180. Cellulolytic actinobacteria are cellulase-producing bacteria capable of degrading cellulose. This study aimed to isolate, characterize, evaluate the cellulolytic ability, and to determine physiological characterization of soil cellulolytic actinobacteria isolated from the Ujung Pancu area, Aceh Besar. Isolation of actinobacteria from soil samples was performed using serial dilution method on Yeast Malt Agar (YMA) medium. Morphological characterization was carried out by growing isolates on YMA, Oatmeal Agar (OA), and Yeast Starch Agar (YSA) media. Cellulolytic ability was determined by calculating the cellulolytic index (IS) on 1% carboxymethyl cellulose (CMC) medium after adding 0.1% congo red solution. Physiological characterization of cellulolytic actinobacteria tested in this study was salinity, pH, and carbon source in liquid Yeast Malt (liquid YM), and the growth was measured at a wavelength of 581nm. The results showed that a total of nine isolates of actinobacteria were isolated, which belonged to the genus Streptomyces. Cellulolytic test results showed that eight isolates had the ability to degrade cellulose. Isolates AUP-04, AUP-03, and AUP-01 had the highest cellulolytic index value. Physiological characterization results revealed that three isolates had different tolerances for salinity levels, pH, and types of carbon sources. AUP-03 isolate grew well at 10% salinity with an OD value of 0.88, isolate AUP-01 grew at 5% salinity with an OD value of 0.49, whereas isolate AUP-04 grew well on media that did not contain salinity. All three isolates grew well at pH 6 with OD values of 0.93, 1.12, and 1.27. AUP-03 and AUP-01 isolates grew well on media containing dextrose as carbon source with OD values of 0.154 and 0.17, respectively, while isolate AUP-04 grew well on glucose-containing media with an OD value of 0.22.

scholarly journals Pichia pastoris as a host system for transformations.

1985 ◽  
Vol 5 (12) ◽  
pp. 3376-3385 ◽  
Author(s):  
J M Cregg ◽  
K J Barringer ◽  
A Y Hessler ◽  
K R Madden
Keyword(s):  
Pichia Pastoris ◽  
Auxotrophic Mutant ◽  
Methylotrophic Yeast ◽  
Fusion Procedure ◽  
Methylotrophic Yeast Pichia Pastoris ◽  
Isolation And Characterization ◽  
Mutant Host ◽  
Histidinol Dehydrogenase ◽  
His4 Gene

We developed a methylotrophic yeast, Pichia pastoris, as a host for DNA transformations. The system is based on an auxotrophic mutant host of P. pastoris which is defective in histidinol dehydrogenase. As a selectable marker, we isolated and characterized the P. pastoris HIS4 gene. Plasmid vectors which contained either the P. pastoris or the Saccharomyces cerevisiae HIS4 gene transformed the P. pastoris mutant host. DNA transfer was accomplished by a modified version of the spheroplast generation (CaCl2-polyethylene glycol)-fusion procedure developed for S. cerevisiae. In addition, we report the isolation and characterization of P. pastoris DNA fragments with autonomous replication sequence activity. Two fragments, PARS1 and PARS2, when present on plasmids increased transformation frequencies to 10(5)/micrograms and maintained the plasmids as autonomous elements in P. pastoris cells.

scholarly journals Isolation and Characterization of a Molybdenum-reducing and Carbamate-degrading Serratia sp. strain Amr-4 in soils from Egypt

2021 ◽  
Vol 3 (2) ◽  
pp. 25-32
Author(s):  
M. Abd. AbdEl-Mongy ◽  
M.F. Rahman ◽  
Mohd Yunus Shukor
Keyword(s):  
Carbon Sources ◽  
Biochemical Investigation ◽  
Isolation And Characterization ◽  
Low Concentrations ◽  
Bacteria Inhibition ◽  
High Concentrations ◽  
Reducing Bacteria ◽  
Bioremediation Agent

Physical or chemical procedures could efficiently remove contaminants including pesticides such as carbamates from high concentrations of toxicants. Bioremediation, on the other hand, is frequently a less expensive option in the long term when used at low concentrations. Isolation of multiple toxicants removing microorganisms is the goal of bioremediation. In this paper we report on the molybdenum reduction of the bacterium and its ability to grow on the carbamates carbofuran and carbaryl as carbon sources. Both the carbamates carbofuran and carbaryl cannot support molybdenum reduction when used as the sole carbon sources. Between pH 6.0 and 6.8 and between 30 and 34 oC, the bacterium is most efficient in converting molybdate to Mo-blue. For molybdate reduction, glucose was shown to be the strongest electron donor, with maltose and sucrose coming in second and third, respectively, and d-mannitol and d-adonitol coming in last. Phosphate concentrations of 2.5 to 7.5 mM and molybdate concentrations of 20 to 30 mM are also needed. Identical to that of a decreased phosphomolybdate, the Mo-blue produced by the new Mo-reducing bacteria has an absorption spectrum similar to prior Mo-reducing bacteria. Inhibition of molybdenum reduction was 73.3, 50.1, 50.1 and 20.7 percent, respectively, by mercury, copper, silver and lead at 2 ppm. The bacterium was tentatively identified as Serratia sp. strain Amr-4 after biochemical investigation. This bacterium's ability to detoxify a variety of toxicants is highly sought after, making it a significant bioremediation agent.

scholarly journals Isolation and characterization of fungi from deteriorated old manuscripts from Banyumas, collection of Library of Universitas Indonesia

2021 ◽  
Vol 948 (1) ◽  
pp. 012031
Author(s):  
W Lintang ◽  
T Susetyo-Salim ◽  
A Oetari ◽  
W Sjamsuridzal
Keyword(s):  
Electron Microscopy ◽  
Fungal Spores ◽  
Carbon Sources ◽  
Morphological Characteristics ◽  
Fungal Growth ◽  
Dry Weight ◽  
Isolation And Characterization ◽  
Fungal Isolates ◽  
Scanning Electron

Abstract Fungi are the main cause of old manuscript deterioration since manuscripts provide carbon source and nutrient for fungal growth. Isolation of fungi from deteriorated old manuscripts from Banyumas was carried out and their morphology, xerophilic, and cellulolytic nature were investigated. Two deteriorated old dluwang manuscripts showed fungal spores, brown spots, and discoloured paper. Based on morphological characteristics, 31 fungal isolates belonged to five genera (Aspergillus Micheli, Cladosporium Link, Curvularia Boedijn, Penicillium Link, Ulocladium Preuss). These genera have been reported from deteriorated old manuscripts from several historical places in Indonesia. Xerophilic character was shown by 90% (28 isolates) as determined by growth in DG18 medium, which indicated the ability to grow in dry substrates such as old manuscripts. Cellulolytic character was shown by 93.5% (29 isolates) as determined by growth in dluwang paper and merang paper, which indicated that the papers were used as carbon sources and substrates. After 30 days-incubation, the dry weight loss of merang paper was 0.28-51.2%. Result from Scanning Electron Microscopy showed that the deterioration of merang paper were caused by the isolates as shown by the presence of fungal structures. These results showed that the fungal isolates were able to deteriorate old manuscripts from Banyumas, Indonesia.

Isolation and Characterization of Poly-(R)-3-hydroxybutyrate Produced by Bacillus thuringiensis TH-01

2021 ◽  
Vol 874 ◽  
pp. 81-87
Author(s):  
Wa Ode Sri Rizki ◽  
Zuhdina Sabiqoh ◽  
Enny Ratnaningsih ◽  
Rukman Hertadi
Keyword(s):  
Bacillus Thuringiensis ◽  
Carbon Sources ◽  
Decomposition Temperature ◽  
Infrared Analysis ◽  
Scanning Calorimetry ◽  
Isolation And Characterization ◽  
Bacterium Strain ◽  
Thermal Stability Of ◽  
Termite Nest

Poly-(R)-3-hydroxybutyrate (PHB) is a biopolymer that can be synthesized by several microorganisms such as bacteria, yeast, and fungi as secondary metabolites. PHB is produced by bacteria in a medium containing a limited amount of key nutrients such as nitrogen, phosphorus, and magnesium but rich in carbon sources. PHB is a biodegradable plastic that has many applications in the medical and industrial fields. This study aimed to isolate and characterize a biopolymer produced by a bacterium strain isolated from a termite nest in India that was identified by 16S rRNA method as Bacillus thuringiensis TH-01. The biopolymer was produced by growing the bacteria in a high medium overnight at 37 °C in a shaking incubator at 150 rpm, and the resulting biopolymer was extracted with a mixture of chloroform–NaOCl (1:1). The efficiency of biopolymer production was about 10.545% ± 26.125%. Fourier transform infrared analysis gave prominent absorption peaks at 3400 cm−1 (stretching of O–H), 2900 cm−1 (stretching of C–H), 1700 cm−1 (stretching of C=O), 1280 cm−1 (symmetric deformation of C–H), and 1050 cm−1 (stretching of C−O), confirming that the biopolymer is PHB. The thermal stability of PHB granules as was determined by thermogravimetry analysis (TGA) and differential scanning calorimetry (DSC) showed that the decomposition temperature and of the polymer were 271.6–310.0 °C and 7.48 J/g respectively, and its crystallinity was about 5.12%.

scholarly journals Isolation and characterization of SGE1: a yeast gene that partially suppresses the gal11 mutation in multiple copies.

Genetics ◽  
1993 ◽  
Vol 134 (3) ◽  
pp. 675-683
Author(s):  
H Amakasu ◽  
Y Suzuki ◽  
M Nishizawa ◽  
T Fukasawa
Keyword(s):  
Carbon Sources ◽  
Northern Analysis ◽  
Wild Type ◽  
Reading Frame ◽  
Recessive Mutations ◽  
Isolation And Characterization ◽  
Multiple Copies ◽  
The Right ◽  
Inducible Genes

Abstract Recessive mutations of GAL11 in Saccharomyces cerevisiae cause pleiotropic defects that include weak fermentation of galactose, alpha-specific sterility and slow growth on nonfermentable carbon sources. Recent experiments suggest that Gal11P functions as a "co-activator" that links transcriptional activators, such as Gal4p and Grf1p/Rap1p/Tuf1p, with the basic transcription machinery. In the present experiments we isolated a gene, SGE1, that suppresses gal11 for growth on ethidium bromide/galactose agar when the gene was present in two or more copies. The other gal11 phenotypes were not suppressed by SGE1 in the multiple-copy state. Multiple copies of SGE1 increased expression of galactose-inducible genes in gal11 yeast, presumably at the level of transcription. When SGE1 was disrupted in wild-type yeast, the expression of galactose-inducible genes decreased to 50-60% of the wild-type level, presumably due to effect on transcription. Complete DNA sequence analysis revealed that SGE1 encodes a predicted protein of 543 amino acids. SGE1-specific mRNA of 1.8 kilonucleotides was detected by Northern analysis along the direction of the open reading frame. The gene mapped near RAD56, at the right end of chromosome 16.

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