Characterization of a partially folded intermediate of papain induced by fluorinated alcohols at low pH

Eight yeast strains (denoted as D1 to D8) were isolated from samples of natural fermented pineapple. Strain D8 showed highest alcoholic production at low pH and special aroma of pineapple has been chosen for further study. Taxonomic characterization of strain D8 using morphological, biochemical and molecular biological studies confirmed that strain D8 belong to Saccharomycetaceae family, Saccharomycetales order and Saccharomyces cerevisiae species. Therefore, we named this strain as Saccharomyces cerevisiae D8 for further study on Brandy production from pineapple. Citation: Hoang Thi Le Thuong, Nguyen Quang Hao, Tran Thi Thuy, 2017. Taxonomic characterization and identification of Saccharomyces cerevisiae D8 for brandy production from pineapple. Tap chi Sinh hoc, 39(4): 474- 482. DOI: 10.15625/0866-7160/v39n4.10864.*Corresponding author: [email protected] Received 5 December 2016, accepted 12 August 2017

Download Full-text

Identification and characterization of a second 4,4′-dibenzamido-2,2′-stilbenedisulphonate (DBDS)-binding site on band 3 and its relationship with the anion/proton co-transport function

Biochemical Journal ◽

10.1042/bj20041211 ◽

2005 ◽

Vol 388 (1) ◽

pp. 343-353 ◽

Cited By ~ 3

Author(s):

James M. SALHANY ◽

Karen S. CORDES ◽

Renee L. SLOAN

Keyword(s):

Binding Site ◽

Proton Transport ◽

Chloride Concentration ◽

Band 3 ◽

Low Ph ◽

Binding Mechanism ◽

Access Channel ◽

Binding Kinetic ◽

Transport Site

Band 3 mediates both electroneutral AE (anion exchange) and APCT (anion/proton co-transport). Protons activate APCT and inhibit AE with the same pK (∼5.0). SDs (stilbenedisulphonates) bind to a primary, high-affinity site on band 3 and inhibit both AE and APCT functions. In this study, we present fluorescence and kinetic evidence showing that lowering the pH activates a second site on band 3, which binds DBDS (4,4′-dibenzamido-2,2′-stilbenedisulphonate) independently of chloride concentration, and that DBDS binding to the second site inhibits the APCT function of band 3. Activation of the second site correlated with loss of chloride binding to the transport site, thus explaining the lack of competition. The kinetics of DBDS binding at the second site could be simulated by a slow-transition, two-state exclusive binding mechanism (R0↔T0+D↔TD↔RD, where D represents DBDS, R0 and T0 represent alternate conformational states at the second DBDS-binding site, and TD and RD are the same two states with ligand DBDS bound), with a calculated overall Kd of 3.9 μM and a T0+D↔TD dissociation constant of 55 nM. DBDS binding to the primary SD site inhibited approx. 94% of the proton transport at low pH (KI=68.5±11.8 nM). DBDS binding to the second site inhibited approx. 68% of the proton transport (KI=7.27±1.27 μM) in a band 3 construct with all primary SD sites blocked through selective cross-linking by bis(sulphosuccinimidyl)suberate. DBDS inhibition of proton transport at the second site could be simulated quantitatively within the context of the slow-transition, two-state exclusive binding mechanism. We conclude that band 3 contains two DBDS-binding sites that can be occupied simultaneously at low pH. The binding kinetic and transport inhibition characteristics of DBDS interaction with the second site suggest that it may be located within a gated access channel leading to the transport site.

Download Full-text

Aromatic and Sensory Characterization of Maturana Blanca Wines Made with Different Technologies

Beverages ◽

10.3390/beverages7010010 ◽

2021 ◽

Vol 7 (1) ◽

pp. 10

Author(s):

Ana Naranjo ◽

Leticia Martínez-Lapuente ◽

Belén Ayestarán ◽

Zenaida Guadalupe ◽

Irene Pérez ◽

...

Keyword(s):

Gas Chromatography ◽

Sensory Analysis ◽

Low Ph ◽

Ethyl Esters ◽

Vitis Vinifera L ◽

White Variety ◽

White Wines ◽

Mass Detector ◽

Oak Barrels

Vitis vinífera L. cv. Maturana Blanca is an autochthonous minor variety recently recovered in the Rioja Qualified Denomination of Origin (D.O.Ca Rioja, Spain) for the production of monovarietal white wines with singular and differentiated characteristics. In this paper, Maturana Blanca wines made with different technologies were analyzed by sensory analysis and aromatic profile by gas chromatography-mass detector. Maturana Blanca wines were characterized by low pH, high acidity, and yellowish tonalities. The compounds that most influenced the aroma of Maturana Blanca wines were those related to fruity (acetates and ethyl esters), floral aromas (2-phenylethanol), and spicy notes (γ-decalactone). These wines were mainly characterized by volatile compounds of fruity aromas of banana and apple. The use of pre-fermentative maceration increased the concentration of ethyl esters and acetates and produced wines with higher odor activity values, indicating a greater aromatic intensity. The aromatic profile of Maturana Blanca wines fermented in oak barrels showed a greater complexity as they were also characterized by the presence of important amounts of furfural, whiskey lactone, and eugenol. The sensory analysis confirmed the results obtained in the aromatic analysis, and described the wines as fresh and balanced in mouth, with notes of acidity and medium to high persistence. These results will contribute to a better knowledge of this white variety.

Download Full-text

Characterization of Lassa Virus Cell Entry and Neutralization with Lassa Virus Pseudoparticles

Journal of Virology ◽

10.1128/jvi.01711-08 ◽

2009 ◽

Vol 83 (7) ◽

pp. 3228-3237 ◽

Cited By ~ 34

Author(s):

François-Loic Cosset ◽

Philippe Marianneau ◽

Geraldine Verney ◽

Fabrice Gallais ◽

Noel Tordo ◽

...

Keyword(s):

Immune Response ◽

Humoral Immune Response ◽

Neutralizing Antibodies ◽

Cell Attachment ◽

Low Ph ◽

Cell Entry ◽

Lassa Virus ◽

Ph Optimum ◽

Humoral Immune

ABSTRACT The cell entry and humoral immune response of the human pathogen Lassa virus (LV), a biosafety level 4 (BSL4) Old World arenavirus, are not well characterized. LV pseudoparticles (LVpp) are a surrogate model system that has been used to decipher factors and routes involved in LV cell entry under BSL2 conditions. Here, we describe LVpp, which are highly infectious, with titers approaching those obtained with pseudoparticles displaying G protein of vesicular stomatitis virus and their the use for the characterization of LV cell entry and neutralization. Upon cell attachment, LVpp utilize endocytic vesicles for cell entry as described for many pH-dependent viruses. However, the fusion of the LV glycoproteins is activated at unusually low pH values, with optimal fusion occurring between pH 4.5 and 3, a pH range at which fusion characteristics of viral glycoproteins have so far remained largely unexplored. Consistent with a shifted pH optimum for fusion activation, we found wild-type LV and LVpp to display a remarkable resistance to exposure to low pH. Finally, LVpp allow the fast and quantifiable detection of neutralizing antibodies in human and animal sera and will thus facilitate the study of the humoral immune response in LV infections.

Download Full-text

Biochemical and functional characterization of Mycobacterium tuberculosis ketol-acid reductoisomerase

Microbiology ◽

10.1099/mic.0.001087 ◽

2021 ◽

Vol 167 (9) ◽

Author(s):

Nirbhay Singh ◽

Anu Chauhan ◽

Ram Kumar ◽

Sudheer Kumar Singh

Keyword(s):

Amino Acids ◽

Mycobacterium Tuberculosis ◽

Type Species ◽

Low Ph ◽

Stress Conditions ◽

Starvation Stress ◽

Content Type ◽

E Coli ◽

Link Type

Branched-chain amino acids (BCAAs) are essential amino acids, but their biosynthetic pathway is absent in mammals. Ketol-acid reductoisomerase (IlvC) is a BCAA biosynthetic enzyme that is coded by Rv3001c in Mycobacterium tuberculosis H37Rv (Mtb-Rv) and MRA_3031 in M. tuberculosis H37Ra (Mtb-Ra). IlvCs are essential in Mtb-Rv as well as in Escherichia coli . Compared to wild-type and IlvC-complemented Mtb-Ra strains, IlvC knockdown strain showed reduced survival at low pH and under low pH+starvation stress conditions. Further, increased expression of IlvC was observed under low pH and starvation stress conditions. Confirmation of a role for IlvC in pH and starvation stress was achieved by developing E. coli BL21(DE3) IlvC knockout, which was defective for growth in M9 minimal medium, but growth could be rescued by isoleucine and valine supplementation. Growth was also restored by complementing with over-expressing constructs of Mtb-Ra and E. coli IlvCs. The E. coli knockout also had a survival deficit at pH=5.5 and 4.5 and was more susceptible to killing at pH=3.0. The biochemical characterization of Mtb-Ra and E. coli IlvCs confirmed that both have NADPH-dependent activity. In conclusion, this study demonstrates the functional complementation of E. coli IlvC by Mtb-Ra IlvC and also suggests that IlvC has a role in tolerance to low pH and starvation stress.

Download Full-text

Structural Characterization of Charcoal Exposed to High and Low Ph: Implications for 14C Sample Preparation and Charcoal Preservation

Radiocarbon ◽

10.1017/s0033822200033592 ◽

2008 ◽

Vol 50 (2) ◽

pp. 289-307 ◽

Cited By ~ 59

Author(s):

N R Rebollo ◽

I Cohen-Ofri ◽

R Popovitz-Biro ◽

O Bar-Yosef ◽

L Meignen ◽

...

Keyword(s):

Alkali Treatment ◽

Low Ph ◽

Upper Paleolithic ◽

Salt Bridges ◽

Weight Losses ◽

The Reformation ◽

Early Upper Paleolithic ◽

Early Upper

Chemical and structural similarities between poorly preserved charcoal and its contaminants, as well as low radiocarbon concentrations in old samples, complicate 14C age determinations. Here, we characterize 4 fossil charcoal samples from the late Middle Paleolithic and early Upper Paleolithic strata of Kebara Cave, Israel, with respect to the structural and chemical changes that occur when they are subjected to the acid-base-acid (ABA) treatment. Differential thermal analysis and TEM show that acid treatment disrupts the structure, whereas alkali treatment results in the reformation of molecular aggregates. The major changes are ascribed to the formation of salt bridges at high pH and the disruption of the graphite-like crystallites at low pH. Weight losses during the treatments are consistently greater for older samples, implying that they are less well preserved. Based on the changes observed in vitro due to pH fluctuations, various methods for removing contamination, as well as a mechanism for preferential preservation of charcoal in nature, are proposed.

Download Full-text

ISOLATION, SCREENING, AND CHARACTERIZATION OF PROBIOTICS (LACTIC ACID BACTERIA) ANTAGONISTIC AGAINST Candida albicans

Metamorfosa Journal of Biological Sciences ◽

10.24843/metamorfosa.2017.v04.i02.p19 ◽

2017 ◽

Vol 4 (2) ◽

pp. 263

Author(s):

Ida Ayu Ketut Ariningsih ◽

Yan Ramona ◽

Nyoman Semadi Antara

Keyword(s):

Candida Albicans ◽

Lactic Acid ◽

Lactic Acid Bacteria ◽

Reproductive Tract ◽

Antagonistic Activity ◽

Low Ph ◽

Production Test ◽

Ph Conditions ◽

High Level

Candidacies in female reproductive tract are mainly caused by Candida albicans. This infection often causes serious problems, particularly on their reproductive tract (genital part). Until recently, control of this infection has relied on the use of antibiotics. However due to numerous bad side effects of antibiotics, lactic acid bacteria have been proposed as an alternative method to control the growth of Candida albicans. Therefore, this research was aimed to isolate, screen, and characterize lactic acid bacterial isolates (LAB) antagonistic against Candida albicans (the causative agent of candidacies infection in reproductive tract of human). LABs were isolated from various fermented foods, such as tape ketan and kimchi. Isolation of LABs was conducted by applying dilution and spread plate method on MRS agar medium supplemented with BCP indicator to distinguish LABs from non acid-producing bacteria. Colonies with indication to produce acid were screened for antagonistic activity against C. albicans on MRS agar and followed by characterization of those isolates (Gram stain, catalase production test, oxydase production, gas production test, resistance test to low pH conditions and to high level of NaDC (sodium deoxicolic), and test for ability to convert colic acid (CA) into deoxicolic acid (DCA)). The results showed that 46 LAB isolates were successfully isolated from samples of tape ketan and kimchi. Among those, 7 isolates showed antagonistic activity against C. albicans in in vitro tests. All these 7 candidates were also found to be resistance to low pH conditions (up to pH 2) and to high level of NaDC (up to 0.6 mM). Four most potential isolates were further testes for ability to convert colic acid into deoxycolic acid and none showed positive result, indicating that they all showed initial potential and safe for future human probiotic development (especially to be used to treat patients infected by C. albicans).

Download Full-text