Hypouricemic, hepatoprotective and nephroprotective roles of oligopeptides derived from Auxis thazard protein in hyperuricemic mice

The extensive use of antibiotics leads to an increasing number of antibiotic-resistant pathogenic microorganisms. The development of new antimicrobials is needed for clinical, veterinary, and food applications. Bacteriocins are small peptides with antimicrobial activity ribosomally synthesized by bacteria and could be applied as an alternative to classical antibiotics. In this study, the bacteriocin-like (BLIS) peptide, produced by Staphylococcus xylosus was partially purified and main characteristics (pH, thermal stability, resistance to some protease enzymes and molecular weight) were evaluated. Ammonium sulfate precipitation, acetone extraction and ion-exchange chromatography methods were applied for purification of bacteriocin. The activity of bacteriocin was detected using a well diffusion assay method and the amount of protein concentration was estimated by Lowry method. Molecular weight (~ 6 kDa) of purified bacteriocin was determined by sodium dodecyl sulphate polyacrilamide gel electrophoresis (SDS PAGE) method. The highest purification yield (80 %) was obtained using ion-exange chromatography and SP-sepharose as sorbent. The purified bacteriocin remained stable at pH values between 2.0 and 12.0 for 4 h. No decrease in antibacterial activity was estimated after 30 min at 121º C temperature. The purified bacteriocin was resistant to papain, pepsin and trypsin action. The BLIS inhibits a growth of Listeria monocytogenes (93 ± 3.0 %), Bacillus subtilis (85 ± 4.0 %), Pediococcus pentosaceus (79 ± 4.0 %), Staphylococcus aureus (51 ± 5.0 %) and Propionibacterium acnes (70 ± 5 %) up to 24 hours. Such bacteriocin preparation could be applied as antimicrobial agent in medical and food industry.

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Protex 51FP as a starter for accelerating fish sauce fermentation from anchovy (Stolephorus commersonii)

Emirates Journal of Food and Agriculture ◽

10.9755/ejfa.2021.v33.i5.2701 ◽

2021 ◽

pp. 379

Author(s):

Le Minh Chau ◽

Ho Thi Bich Ngoc ◽

Claire Donnay Moreno ◽

Sandrine Bruzac ◽

Jean-Pascal Bergé ◽

...

Keyword(s):

Amino Acids ◽

Molecular Weight ◽

Starter Culture ◽

Essential Amino Acids ◽

Control Group ◽

Fish Sauce ◽

Small Peptides ◽

Traditional Methods ◽

Commercial Fish ◽

Salt Addition

In this 180-day study, commercial Protex 51FP enzyme effects as a starter culture on anchovy fish sauce fermentation were investigated. Three fish source fermentation groups, including a control group (the anchovy with 25% of salt addition), E group (the anchovy with 25% of salt and 1% of Protex 51FP addition), and E (-s) group (the anchovy with 1% of Protex 51FP and after 6 hours with 25% of salt), were compared. The fish sauce fermentation groups were sampled, packed into glass jars (10 liters), and covered by a lid at ambient temperature (22 - 30°C) for 180 days. Three commercial fish sauces were also included as nutritional references. The results showed that the addition of Protex 51FP achieved positive results of total nitrogen content and amino acids compared to the control samples (p<0.05). These values were competitive with commercial product figures. Total amino acids in 8000mg/100ml fish sauce were significantly higher than those in the control. There were rich in essential amino acids (41-43%) and small peptides (13% peptides with a molecular weight below 200 Da, 32-39% peptides with a molecular weight below 130 - 200 Da, and 25-28% of peptides with a molecular weight below 200 – 360 Da). Compared with traditional methods, the addition of Protex 51FP (p<0.05) could improve the quality of fish sauce and obtain greater nutritional values. In all experiments, the color of adding-enzyme samples was darker than that of the traditional products, and the smell of these samples (including traditional methods) was not as quite strong as commercial products.

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Dihydrofolate reductase: low-resolution mass-spectrometric analysis of an elastase digest as a sequencing tool

Biochemical Journal ◽

10.1042/bj1370409 ◽

1974 ◽

Vol 137 (2) ◽

pp. 409-411 ◽

Cited By ~ 30

Author(s):

Howard R. Morris ◽

Karen E. Batley ◽

Nigel G. L. Harding ◽

Richard A. Bjur ◽

John G. Dann ◽

...

Keyword(s):

Mass Spectrometry ◽

Molecular Weight ◽

Dihydrofolate Reductase ◽

Mass Spectrometric ◽

Spectrometric Analysis ◽

Small Peptides ◽

Molecular Weight Range ◽

Unknown Structure ◽

Resolution Mass ◽

The Ideal

An elastase digest of a protein of unknown structure, dihydrofolate reductase, was studied by mass spectrometry. This soluble digest contained a large number of small peptides in different yields, within the ideal molecular-weight range (200–1200) for mixture-analysis mass spectrometry. Sequences of the major component peptides in the digest are reported.

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Recent Advances in Small Peptides of Marine Origin in Cancer Therapy

Marine Drugs ◽

10.3390/md19020115 ◽

2021 ◽

Vol 19 (2) ◽

pp. 115

Author(s):

Qi-Ting Zhang ◽

Ze-Dong Liu ◽

Ze Wang ◽

Tao Wang ◽

Nan Wang ◽

...

Keyword(s):

Molecular Weight ◽

Causes Of Death ◽

Biological Species ◽

The Body ◽

Small Peptides ◽

Small Molecular Weight ◽

Peptide Bonds ◽

Preclinical Treatment ◽

Fast Absorption ◽

Proteins And Peptides

Cancer is one of the leading causes of death in the world, and antineoplastic drug research continues to be a major field in medicine development. The marine milieu has thousands of biological species that are a valuable source of novel functional proteins and peptides, which have been used in the treatment of many diseases, including cancer. In contrast with proteins and polypeptides, small peptides (with a molecular weight of less than 1000 Da) have overwhelming advantages, such as preferential and fast absorption, which can decrease the burden on human gastrointestinal function. Besides, these peptides are only connected by a few peptide bonds, and their small molecular weight makes it easy to modify and synthesize them. Specifically, small peptides can deliver nutrients and drugs to cells and tissues in the body. These characteristics make them stand out in relation to targeted drug therapy. Nowadays, the anticancer mechanisms of the small marine peptides are still largely not well understood; however, several marine peptides have been applied in preclinical treatment. This paper highlights the anticancer linear and cyclic small peptides in marine resources and presents a review of peptides and the derivatives and their mechanisms.

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Simplified isolation and molecular composition of NADH dehydrogenase of the respiratory chain

Biochemical Journal ◽

10.1042/bj2030477 ◽

1982 ◽

Vol 203 (2) ◽

pp. 477-481 ◽

Cited By ~ 17

Author(s):

C Paech ◽

A Friend ◽

T P Singer

Keyword(s):

Molecular Weight ◽

Nadh Dehydrogenase ◽

Low Molecular Weight ◽

Stable Form ◽

Small Peptides ◽

Rapid Preparation ◽

Binding Peptides ◽

Simplified Procedure ◽

Haem Iron

A simplified procedure for the isolation of NADH dehydrogenase from the inner membrane of ox heart mitochondria is presented which permits relatively rapid preparation of the enzyme in a more stable form than that afforded by published methods. The protein thus isolated displays more than eight different subunits in gel electrophoresis under denaturing conditions, three of which are also present in the "low-molecular-weight form' of the enzyme prepared under more drastic conditions. Complex I contains several subunits, mostly of low molecular weight, not seen in soluble purified NADH dehydrogenase. It is suggested that some of these may be ‘binding peptides’ necessary in linking NADH dehydrogenase to ubiquinone reduction, analogously to the role of small peptides in linking succinate dehydrogenase to ubiquinone. The dehydrogenase isolated by the rapid method contains equimolar amounts of non-haem iron and labile sulphur, but on further manipulation non-haem iron (but no labile sulphur) is lost, resulting in ratios of S/Fe in excess of unity, as previously reported for preparations isolated by longer procedures.

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Unique Degradation Of Fibrinogen By Western Diamondback Rattlesnake Venom Proteases

10.1055/s-0038-1652104 ◽

1981 ◽

Author(s):

Bharat V Pandya ◽

Andrei Z Budzynski ◽

Ronald N Rubin ◽

Stephanie A Olexa

Keyword(s):

Molecular Weight ◽

Gel Electrophoresis ◽

Polyacrylamide Gel Electrophoresis ◽

Fibrin Clot ◽

Polyacrylamide Gel ◽

Small Peptides ◽

Crude Venom ◽

Degradation Pattern ◽

Diamondback Rattlesnake ◽

Human Plasminogen

The venom of western diamondback rattlesnake, Crotalus atrox, renders blood unclottable even after addition of thrombin. The phenomenon is due to a limited proteolytic degradation of fibrinogen. Purified fibrinogen (1 mg/ml) or human plasma incubated with crude venom (7.5 μg/ml) became unclottable. Fibrinogen in a purified system was affected at a faster rate than that in plasma suggesting that plasma protease inhibitors partially inhibited the proteolysis of fibrinogen. The degradation of plasma fibrinogen did not result from plasminogen activation since the venom did not activate purified human plasminogen as determined by an amidolytic assay and SDS polyacrylamide gel electrophoresis. The degradation of the fibrinogen molecule (Mr 340,000) proceeded steadily with a continuous cleavage of small peptides as demonstrated by SDS polyacrylamide gel electrophoresis. The bulk of the degraded fibrinogen was represented in gels as a single band with gradually increasing electrophoretic mobility as the incubation time progressed. The correlation of molecular weight and clottability demonstrated that the earliest unclottable derivative had Mr 285,000; its Aα and Bβ chains were degraded loosing peptides of Mr 20,000 and 6,500, respectively, but the γ chain appeared intact. The degradation pattern of fibrinogen in plasma was different than that of fibrinogen alone. An unclottable fibrinogen derivative isolated from plasma by precipitation with 2.2 M glycine had Mr 325,000; its Aα and γ chains appeared unaffected but the Bβ chain had lost a peptide of Mr 6,500. This product represents a novel unclottable derivative of fibrinogen with apparently intact Aα and γ chains, cleaved Bβ chain and high molecular weight. The cleavage of the Bβ chain would indicate that the intact Bβ chain may have an important role in the conversion of fibrinogen to fibrin clot.

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In vitro availability of iron and zinc: effects of the type, concentration and fractions of digestion products of the protein

British Journal Of Nutrition ◽

10.1079/bjn19960079 ◽

1996 ◽

Vol 76 (5) ◽

pp. 727-741 ◽

Cited By ~ 11

Author(s):

Francisca Pérez-Llamas ◽

Mechteldis G. E. Diepenmaat-Wolters ◽

Salvador Zamora

Keyword(s):

Molecular Weight ◽

Free Amino ◽

Essential Element ◽

Lower Percentage ◽

Small Peptides ◽

Digestion Process ◽

Iron And Zinc ◽

Dialysis Method ◽

Positive Effect

An in vitro dialysis method was employed to determine the effect on the Fe and Zn absorption of the type (beef, pork and soyabean) and the amount (10 and 30 g/kg) of protein present. In addition, the effects of low- and high-molecular-weight (LMW and HMW respectively) digestion products were investigated. After in vitro digestion and dialysis a lower percentage of N, Fe and Zn was found in the LMW fractions from beef, pork and soyabean proteins when the protein level was increased from 10 to 30 g/kg; the higher level of protein being associated with a lower percentage of hydrolysed protein. The highest percentage levels of intrinsic Fe were always found in the HMW fractions, independent of the type and the level of proteins studied, while in the case of Zn, both HMW and LMW fractions gave similar values. An interaction was found between inorganic Zn and non-haem-Fe. The addition of inorganic Zn (10 μg/ml) caused a significant decrease in the in vitro availability of Fe from soyabean protein, while it did not affect the dialysability of intrinsic Fe from beef and pork proteins. Our results showed that the type and the level of the protein had a positive effect on the dialysability of extrinsic Fe. We postulate that the effect of a protein on the absorption of extrinsic Fe could be accounted for by free amino acids and/or small peptides released during the digestion process and also by the undigested or partially-digested HMW fractions of hydrolysed proteins which could play a fundamental role in the availability of this essential element.

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Evaluation of the dark field method for large association of spread DNA

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100081814 ◽

1978 ◽

Vol 36 (2) ◽

pp. 190-191

Author(s):

Douglas C. Barker

Keyword(s):

Electron Microscopy ◽

Molecular Weight ◽

Mitochondrial Dna ◽

Extraction Method ◽

Field Method ◽

Dark Field ◽

Kinetoplast Dna ◽

Field Electron ◽

Field Electron Microscopy ◽

Dark Field Electron

A number of satisfactory methods are available for the electron microscopy of nicleic acids. These methods concentrated on fragments of nuclear, viral and mitochondrial DNA less than 50 megadaltons, on denaturation and heteroduplex mapping (Davies et al 1971) or on the interaction between proteins and DNA (Brack and Delain 1975). Less attention has been paid to the experimental criteria necessary for spreading and visualisation by dark field electron microscopy of large intact issociations of DNA. This communication will report on those criteria in relation to the ultrastructure of the (approx. 1 x 10-14g) DNA component of the kinetoplast from Trypanosomes. An extraction method has been developed to eliminate native endonucleases and nuclear contamination and to isolate the kinetoplast DNA (KDNA) as a compact network of high molecular weight. In collaboration with Dr. Ch. Brack (Basel [nstitute of Immunology), we studied the conditions necessary to prepare this KDNA Tor dark field electron microscopy using the microdrop spreading technique.

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Introduction

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100070266 ◽

1978 ◽

Vol 36 (3) ◽

pp. 543-544

Author(s):

W. Bernard

Keyword(s):

Molecular Weight ◽

Electron Microscope ◽

Nucleic Acid ◽

Gene Mapping ◽

Molecular Genetics ◽

Cell Biology ◽

Gene Activity ◽

Close Connection ◽

Basic Information ◽

Simple Systems

In comparison to many other fields of ultrastructural research in Cell Biology, the successful exploration of genes and gene activity with the electron microscope in higher organisms is a late conquest. Nucleic acid molecules of Prokaryotes could be successfully visualized already since the early sixties, thanks to the Kleinschmidt spreading technique - and much basic information was obtained concerning the shape, length, molecular weight of viral, mitochondrial and chloroplast nucleic acid. Later, additonal methods revealed denaturation profiles, distinction between single and double strandedness and the use of heteroduplexes-led to gene mapping of relatively simple systems carried out in close connection with other methods of molecular genetics.

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The External Shape of Human γ GI Immunoglobulin from Crystal Sections

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010006619x ◽

1971 ◽

Vol 29 ◽

pp. 428-429

Author(s):

L. W. Labaw

Keyword(s):

Molecular Weight ◽

Sodium Phosphate ◽

Osmium Tetroxide ◽

Unit Cell ◽

Monoclinic Space Group ◽

X Ray ◽

Large Molecular Weight ◽

Cell Dimensions ◽

Unit Cell Dimensions ◽

Crystallographic Axes

Crystals of a human γGl immunoglobulin have the external morphology of diamond shaped prisms. X-ray studies have shown them to be monoclinic, space group C2, with 2 molecules per unit cell. The unit cell dimensions are a = 194.1, b = 91.7, c = 51.6Å, 8 = 102°. The relatively large molecular weight of 151,000 and these unit cell dimensions made this a promising crystal to study in the EM.Crystals similar to those used in the x-ray studies were fixed at 5°C for three weeks in a solution of mother liquor containing 5 x 10-5M sodium phosphate, pH 7.0, and 0.03% glutaraldehyde. They were postfixed with 1% osmium tetroxide for 15 min. and embedded in Maraglas the usual way. Sections were cut perpendicular to the three crystallographic axes. Such a section cut with its plane perpendicular to the z direction is shown in Fig. 1.This projection of the crystal in the z direction shows periodicities in at least four different directions but these are only seen clearly by sighting obliquely along the micrograph.

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