scholarly journals GENETICS OF EXTRACELLULAR PROTEASE PRODUCTION IN SACCHAROMYCOPSIS LIPOLYTICA

Genetics ◽  
1977 ◽  
Vol 87 (4) ◽  
pp. 621-632
Author(s):  
David M Ogrydziak ◽  
Robert K Mortimer
Keyword(s):  
Extracellular Enzymes ◽  
Skim Milk ◽  
Extracellular Enzyme ◽  
Extracellular Protease ◽  
Protease Production ◽  
Mating Frequency ◽  
Wild Type ◽  
Saccharomycopsis Lipolytica ◽  
Complementation Groups ◽  
Membrane Components

ABSTRACT Mutants of Saccharomycopsis lipolytica with reduced ability to produce zones of clearing on skim-milk agar plates were isolated and their properties studied. For 18 mutants it was possible to score unambiguously segregants of crosses between these mutants and wild type for extracellular protease production. These mutants all produce reduced levels of extracellular protease in liquid culture. The mutations are recessive and are in nuclear genes. The 18 mutations define 10 or 11 complementation groups, no two of which are closely linked. Mutants in four of the complementation groups also produced reduced levels of extracellular RNAse, and the reduced levels of extracellular protease and RNAse production segregate together. Five of the mutants exhibited reduced mating frequency, and one mutant was osmotic remedial for extracellular protease production. These results demonstrate that many genes can affect extracellular protease production. Besides mutations in the structural gene and in regulatory genes, mutations are likely to be in genes involved in steps common to the production of several extracellular enzymes or in genes coding for cell wall or membrane components necessary for extracellular enzyme production.

scholarly journals Extracellular Protease and DNase Activities in Clinical and Environmental Isolates of Cryptococcus neoformans Species Complex from Central India

2019 ◽  
Vol 9 (4-A) ◽  
pp. 328-333
Author(s):  
Richa Gumasta ◽  
Shesh Rao Nawange ◽  
Shankar Mohan Singh ◽  
Abhijeet Garg ◽  
Ruchi Sethi ◽  
...  
Keyword(s):  
Cryptococcus Neoformans ◽  
Species Complex ◽  
Extracellular Enzymes ◽  
Central India ◽  
Vital Role ◽  
Extracellular Protease ◽  
Protease Production ◽  
Environmental Isolates ◽  
Clinical Strains ◽  
Significant Difference

Enzymes are important not only for the growth and multiplication of the microorganism but also in the infection, penetration of the host tissue and encountering host defense mechanisms. This study aims to investigate extracellular protease and DNase activity in clinical (20) and 120 environmental isolates of C. neoformans species complex collected from different localities of central India.  DNase test agar containing toluidine blue and Yeast Carbon Base (YCB) agar medium supplemented with 0.1% BSA + 0.01% polypeptone was employed for the screening of DNase and protease production respectively. DNase and protease production was detected by the appearance of clear zones around the colonies. On the basis of enzymatic activity and their Pz values, high protease production (Pz≤0.6) was observed by 14 (11.6 %) environmental and 4 (11.6 %) clinical strains on 5th day, whereas 35 (29.16 %) environmental and 8 (40 %) clinical strains were screened on 8th day of incubation. Similarly 13 (10.83%) environmental and 3 (15 %) clinical strains on the 5th day, however 32 (26.66%) environmental and 8 (40 %) clinical strains on the 8th day of incubation were found to be high DNase producing strains with low Pz value (Pz≤0.6). In the case of protease activity, no significant difference was observed whereas a significant difference has shown by clinical C. neoformans and C. gattii strains on the 5th day of DNase production (p < .001). Extracellular enzymes play a vital role in the pathogenicity and virulence of C. neoformans species complex, therefore, enzymes are considered as worthy targets for developing therapeutics. Keywords: Cryptococcus neoformans species complex, Extracellular enzymes, DNase, protease, virulence, Pz value

Extracellular Protease, Extracellular Haemolysin, and Virulence in Aeromonas salmonicida

1984 ◽  
Vol 41 (9) ◽  
pp. 1354-1360 ◽  
Author(s):  
Jeremy L. Hackett ◽  
William H. Lynch ◽  
William D. Paterson ◽  
David H. Coombs
Keyword(s):  
Ethidium Bromide ◽  
Enzyme Activities ◽  
Extracellular Enzymes ◽  
Virulent Strain ◽  
Extracellular Enzyme ◽  
Aeromonas Salmonicida ◽  
Extracellular Protease ◽  
Acute Form ◽  
Simultaneous Loss ◽  
Different Strains

We have examined the possibile relationships between extracellular protease and haemolysin and virulence in Aeromonas salmonicida, and the possible plasmid-encoded origin of these extracellular enzyme activities. Variants, isolated from different strains, showed simultaneous loss of both protease and haemolysin. The frequency of spontaneous loss from one strain (0.1–0.4%) as well as the frequency of induced loss from four strains (0.3–18%) treated with ethidium bromide suggested a plasmid-encoded origin for these enzymes. However, plasmid analyses showed no detectable loss of plasmids correlated with the loss of the extracellular activities. There was no change in the LD100 when fish were exposed to one virulent strain or its protease, haemolysin-negative variants indicating that these extracellular activities are not necessary for virulence or pathogenicity in the acute form of furunculosis. Furthermore, both protease, haemolysin-negative and protease, haemolysin-positive clones isolated from a second virulent strain treated with ethidium bromide were avirulent (LD50 increased >4 logs). Thus, attenuation of A. salmonicida can occur without detected loss of the extracellular enzymes, plasmids, or the A-layer.

Cytotoxic Effect of Pseudomonas aurogenosa Protease on Cancer Cells

2019 ◽  
Vol 10 (03) ◽  
Author(s):  
Ghanyia J. Shanyoor ◽  
Fatima R. Abdul ◽  
Nehad A. Taher ◽  
Ihsan A. Raheem
Keyword(s):  
Cell Line ◽  
Normal Cell ◽  
Cytotoxic Effect ◽  
Human Cancer ◽  
Skim Milk ◽  
Exchange Chromatography ◽  
Protease Production ◽  
Normal Cell Line ◽  
Protease Enzyme ◽  
Embryonic Fibroblast

About (20) Pseudomonas rogenosa isolate were experienced for their ability of protease production by calculating the diameter of lysis area after developing on skim milk agar medium (qualitatively ) and the results exhibited that only isolate no (5), was higher isolate for protease making of (26mm) of lysis area. Then, the protein concentration also identified by Bradford method and it was found of 0.16 mg/ ml , then purification was done by using an ion- exchange chromatography with DEAE sephadex G- 100 column and the results showed the presence of 1 peak of enzyme with 50 Kd of molecular weight 2 peaks of other proteins . we tried to investigate the invitro Cytotoxic effect of purified enzyme against two human cancer lines, HeP2 (Human larynx epidermed carcinoma ) , RD ( Rabdo- Sarcoma ) , and one normal cell line Ref ( Rat embryonic fibroblast ) . The cancer and normal cells were treated with different concentrations of protease enzyme ranging from ( 0.05, 0.1, 0.2, 0.4,0.8and 0.16 mg/ml) then incubated for additional 48h at 37C0 and the results showed highest toxicity ( 80.28%) of protease enzyme on RD , moderate cytotoxicity (45.52%) on Hep andslight toxicity ( 37.12% ) on normal cell line (Ref) in a concentration (0.8mg/ml).

Constraints in Adoption ofModern Farm Machines by Tribal Farmers in Ramgarh District of Jharkhand

2019 ◽  
Vol 6 (03) ◽  
Author(s):  
PK SUNDARAM ◽  
BIKASH SARKAR ◽  
UJJWAL KUMAR ◽  
AP ANURAG ◽  
DK RAGHAV ◽  
...  
Keyword(s):  
Cell Line ◽  
Normal Cell ◽  
Human Cancer ◽  
Skim Milk ◽  
Exchange Chromatography ◽  
Protease Production ◽  
Normal Cell Line ◽  
Protease Enzyme ◽  
Tribal Farmers ◽  
Embryonic Fibroblast

About (20) Pseudomonas rogenosa isolate were experienced for their ability of protease production by calculating the diameter of lysis area after developing on skim milk agar medium (qualitatively ) andamp; the results exhibited that only isolate no (5), was higher isolate for protease making of (26mm) of lysis area. Then, the protein concentration also identified by Bradford method andamp; it was found of 0.16 mg/ ml , then purification was done by using an ion- exchange chromatography with DEAE sephadex G- 100 column andamp; the results showed the presence of 1 peak of enzyme with 50 Kd of molecular weight 2 peaks of other proteins . we tried to investigate the invitro Cytotoxic effect of purified enzyme against two human cancer lines, HeP2 (Human larynx epidermed carcinoma ) , RD ( Rabdo- Sarcoma ) , andamp; one normal cell line Ref ( Rat embryonic fibroblast ). The cancer andamp; normal cells were treated with different concentrations of protease enzyme ranging from ( 0.05, 0.1, 0.2, 0.4,0.8andamp; 0.16 mg/ml) then incubated for additional 48h at 37C 0 andamp; the results showed highest toxicity ( 80.28%) of protease enzyme on RD , moderate cytotoxicity (45.52%) on Hep andamp;slight toxicity ( 37.12% )on normal cell line (Ref) in a concentration (0.8mg/ml).

scholarly journals Extracellular hydrolytic enzyme production by proteolytic bacteria from the Antarctic

2013 ◽  
Vol 34 (3) ◽  
pp. 253-267 ◽  
Author(s):  
Mauro Tropeano ◽  
Susana Vázquez ◽  
Silvia Coria ◽  
Adrián Turjanski ◽  
Daniel Cicero ◽  
...  
Keyword(s):  
Extracellular Enzymes ◽  
Restriction Analysis ◽  
Hydrolytic Enzyme ◽  
Hydrolytic Enzymes ◽  
Extracellular Enzyme ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Potter Cove ◽  
Starting Point ◽  
Dna Restriction

AbstractCold−adapted marine bacteria producing extracellular hydrolytic enzymes are important for their industrial application and play a key role in degradation of particulate organic matter in their natural environment. In this work, members of a previously−obtained protease−producing bacterial collection isolated from different marine sources from Potter Cove (King George Island, South Shetlands) were taxonomically identified and screened for their ability to produce other economically relevant enzymes. Eighty−eight proteolytic bacterial isolates were grouped into 25 phylotypes based on their Amplified Ribosomal DNA Restriction Analysis profiles. The sequencing of the 16S rRNA genes from representative isolates of the phylotypes showed that the predominant culturable protease−producing bacteria belonged to the class Gammaproteobacteria and were affiliated to the genera Pseudomonas, Shewanella, Colwellia, and Pseudoalteromonas, the latter being the predominant group (64% of isolates). In addition, members of the classes Actinobacteria, Bacilli and Flavobacteria were found. Among the 88 isolates screened we detected producers of amylases (21), pectinases (67), cellulases (53), CM−cellulases (68), xylanases (55) and agarases (57). More than 85% of the isolates showed at least one of the extracellular enzymatic activities tested, with some of them producing up to six extracellular enzymes. Our results confirmed that using selective conditions to isolate producers of one extracellular enzyme activity increases the probability of recovering bacteria that will also produce additional extracellular enzymes. This finding establishes a starting point for future programs oriented to the prospecting for biomolecules in Antarctica.

scholarly journals Response of <i>Nodularia spumigena</i> to <i>p</i>CO<sub>2</sub> – Part 2: Exudation and extracellular enzyme activities

2013 ◽  
Vol 10 (1) ◽  
pp. 567-582 ◽  
Author(s):  
S. Endres ◽  
J. Unger ◽  
N. Wannicke ◽  
M. Nausch ◽  
M. Voss ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Organic Matter ◽  
Enzyme Activities ◽  
Extracellular Enzymes ◽  
Extracellular Enzyme ◽  
High Pco2 ◽  
Nodularia Spumigena ◽  
Extracellular Enzyme Activities ◽  
Pco2 Treatment ◽  
Co2 Concentrations

Abstract. The filamentous and diazotrophic cyanobacterium Nodularia spumigena plays a major role in the productivity of the Baltic Sea as it forms extensive blooms regularly. Under phosphorus limiting conditions Nodularia spumigena have a high enzyme affinity for dissolved organic phosphorus (DOP) by production and release of alkaline phosphatase. Additionally, they are able to degrade proteinaceous compounds by expressing the extracellular enzyme leucine aminopeptidase. As atmospheric CO2 concentrations are increasing, we expect marine phytoplankton to experience changes in several environmental parameters, including pH, temperature, and nutrient availability. The aim of this study was to investigate the combined effect of CO2-induced changes in seawater carbonate chemistry and of phosphate deficiency on the exudation of organic matter, and its subsequent recycling by extracellular enzymes in a Nodularia spumigena culture. Batch cultures of Nodularia spumigena were grown for 15 days under aeration with low (180 μatm), medium (380 μatm), and high (780 μatm) CO2 concentrations. Obtained pCO2 levels in the treatments were on median 315, 353, and 548 μatm CO2, respectively. Extracellular enzyme activities as well as changes in organic and inorganic compound concentrations were monitored. CO2 treatment–related effects were identified for cyanobacterial growth, which in turn influenced the concentration of mucinous substances and the recycling of organic matter by extracellular enzymes. Biomass production was increased by 56.5% and 90.7% in the medium and high pCO2 treatment, respectively, compared to the low pCO2 treatment. In total, significantly more mucinous substances accumulated in the high pCO2 treatment, reaching 363 μg Xeq L−1 compared to 269 μg Xeq L−1 in the low pCO2 treatment. However, cell-specific rates did not change. After phosphate depletion, the acquisition of P from DOP by alkaline phosphatase was significantly enhanced. Alkaline phosphatase activities were increased by factor 1.64 and 2.25, respectively, in the medium and high compared to the low pCO2 treatment. We hypothesise from our results that Nodularia spumigena can grow faster under elevated pCO2 by enhancing the recycling of organic matter to acquire nutrients.

Wool-degrading Bacillus isolates: extracellular protease production for microbial processing of fabrics

2009 ◽  
Vol 26 (6) ◽  
pp. 1047-1052 ◽  
Author(s):  
Inés Infante ◽  
Maria A. Morel ◽  
Martha C. Ubalde ◽  
Cecilia Martínez-Rosales ◽  
Silvia Belvisi ◽  
...  
Keyword(s):  
Extracellular Protease ◽  
Protease Production

scholarly journals Continuous in vivo infusion of interferon-gamma (IFN-γ) enhances engraftment of syngeneic wild-type cells in Fanca–/– and Fancg–/– mice

Blood ◽  
2006 ◽  
Vol 108 (13) ◽  
pp. 4283-4287 ◽  
Author(s):  
Yue Si ◽  
Samantha Ciccone ◽  
Feng-Chun Yang ◽  
Jin Yuan ◽  
Daisy Zeng ◽  
...  
Keyword(s):  
Stem Cells ◽  
Interferon Gamma ◽  
Cancer Susceptibility ◽  
Genetic Disorder ◽  
Wild Type ◽  
Regulatory Cytokine ◽  
Complementation Groups ◽  
Ifn Γ

Abstract Fanconi anemia (FA) is a heterogeneous genetic disorder characterized by bone marrow (BM) failure and cancer susceptibility. Identification of the cDNAs of FA complementation types allows the potential of using gene transfer technology to introduce functional cDNAs as transgenes into autologous stem cells and provide a cure for the BM failure in FA patients. However, strategies to enhance the mobilization, transduction, and engraftment of exogenous stem cells are required to optimize efficacy prior to widespread clinical use. Hypersensitivity of Fancc–/– cells to interferon-gamma (IFN-γ), a nongenotoxic immune-regulatory cytokine, enhances engraftment of syngeneic wild-type (WT) cells in Fancc–/– mice. However, whether this phenotype is of broad relevance in other FA complementation groups is unresolved. Here we show that primitive and mature myeloid progenitors in Fanca–/– and Fancg–/– mice are hypersensitive to IFN-γ and that in vivo infusion of IFN-γ at clinically relevant concentrations was sufficient to allow consistent long-term engraftment of isogenic WT repopulating stem cells. Given that FANCA, FANCC, and FANCG complementation groups account for more than 90% of all FA patients, these data provide evidence that IFN-γ conditioning may be a useful nongenotoxic strategy for myelopreparation in FA patients.

Production and characterisation of thermostable alkaline protease from Bacillus subtilis isolated from LA hot spring, Terengganu

2021 ◽  
Vol 16 (7) ◽  
pp. 84-91
Author(s):  
Maslinda Alias ◽  
Hakim Che Harun Mohammad ◽  
Ashraf Razali Nurul ◽  
Jasnizat Saidin ◽  
Nazaitulshila Rasit ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Alkaline Protease ◽  
Protease Activity ◽  
Hot Spring ◽  
Skim Milk ◽  
Industrial Applications ◽  
Protease Production ◽  
Significant Activity ◽  
Original Activity ◽  
Optimum Ph

This research aims to produce thermostable alkaline protease from Bacillus subtilis isolated from La Hot Spring, Terengganu, Malaysia. The study was also conducted to determine the optimum conditions for protease production and stability by considering several parameters including pH, temperature and salt concentration. All seven bacteria were screened on skim milk agar overnight at 37 °C. Three strains with the highest proteolytic activity were identified in protease specific medium. The thermostable alkaline protease had an optimum temperature of 60 °C which achieved 85.73, 82.90 and 83.05 U/mL of protease activity for the three strains respectively. Furthermore, the strains exhibited significant activity of more than 90% from their original activity. Meanwhile, the optimum pH for protease production was pH 9 with the protease activity of 76.76, 79.71 and 88.39 U/mL for TB4, TB6 and TB9 strains, respectively. Proteases were found stable at pH 9 where the loss did not exceed 30% of its original activity. Collectively, all of the data emphasised that proteases from B. subtilis were alkaline thermostable proteases in accordance with a recent report. The finding highlights the viability of the proteases for biotechnological and industrial applications.

Temperature Sensitivity of Intracellular and Extracellular Soil Enzyme Activities

2021 ◽  
Author(s):  
Adetunji Alex Adekanmbi ◽  
Laurence Dale ◽  
Liz Shaw ◽  
Tom Sizmur
Keyword(s):  
Enzyme Activity ◽  
Soil Temperature ◽  
Temperature Sensitivity ◽  
Extracellular Enzymes ◽  
Extracellular Enzyme ◽  
Daily Maximum ◽  
Temperature Sensitive ◽  
Intracellular Enzyme ◽  
Temperature Changes ◽  
Som Decomposition

&lt;p&gt;Predicting the pattern of soil organic matter (SOM) decomposition as a feedback to climate change, via release of CO&lt;sub&gt;2&lt;/sub&gt;, is extremely complex and has received much attention. However, investigations often do not differentiate between the extracellular and intracellular processes involved and work is needed to identify their relative temperature sensitivities. Samples were collected from a grassland soil at Sonning, UK with average daily maximum and minimum soil temperature of 15 &amp;#176;C and 5 &amp;#176;C. We measured potential activities of &amp;#946;-glucosidase (BG) and chitinase (NAG) (extracellular enzymes) and glucose-induced CO&lt;sub&gt;2 &lt;/sub&gt;respiration (intracellular enzymes) at a range of assay temperatures (5 &amp;#176;C, 15 &amp;#176;C, 26 &amp;#176;C, 37&lt;sup&gt; &amp;#160;&lt;/sup&gt;&amp;#176;C, and 45 &amp;#176;C). The temperature coefficient Q&lt;sub&gt;10&lt;/sub&gt; (the increase in enzyme activity that occurs after a 10 &amp;#176;C increase in soil temperature) was calculated to assess the temperature sensitivity of intracellular and extracellular enzymes activities. Between 5 &amp;#176;C and 15 &amp;#176;C intracellular and extracellular enzyme activities had equal temperature sensitivity, but between 15 &amp;#176;C and 26&amp;#176;C intracellular enzyme activity was more temperature sensitive than extracellular enzyme activity and between 26 &amp;#176;C and 37 &amp;#176;C extracellular enzyme activity was more temperature sensitive than intracellular enzyme activity. This result implies that extracellular depolymerisation of higher molecular weight organic compounds is more sensitive to temperature changes at higher temperatures (e.g. changes to daily maximum summer temperature) but the intracellular respiration of the generated monomers is more sensitive to temperature changes at moderate temperatures (e.g. changes to daily mean summer temperature). We therefore conclude that the extracellular and intracellular steps of SOM mineralisation are not equally sensitive to changes in soil temperature. The finding is important because we have observed greater increases in average daily minimum temperatures than average daily mean or maximum temperatures due to increased cloud cover and sulphate aerosol emission. Accounting for this asymmetrical global warming may reduce the importance of extracellular depolymerisation and increase the importance of intracellular catalytic activities as the rate limiting step of SOM decomposition.&lt;/p&gt;

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