Genes Encoding Proteolytic Enzymes Fungalysin and Subtilisin in Dermatophytes of Human and Animal Origin: A Comparative Study

2019 ◽  
Author(s):  
Engin Kaplan ◽  
Serpil Gonca ◽  
Hazal Kandemir ◽  
Aylin Döğen ◽  
Süleyha Hilmioğlu-Polat ◽  
...  
Keyword(s):  
Comparative Study ◽  
Proteolytic Enzymes ◽  
Animal Origin ◽  
Genes Encoding

COMPARATIVE ASSESSMENT OF MILK PROTEIN COAGULANTS OF DIFFERENT ORIGIN

2021 ◽  
Author(s):  
А.Г. КРУЧИНИН ◽  
С.Н. ТУРОВСКАЯ ◽  
Е.Е. ИЛЛАРИОНОВА ◽  
А.В. БИГАЕВА
Keyword(s):  
Dairy Products ◽  
Milk Protein ◽  
Proteolytic Enzymes ◽  
Raw Materials ◽  
Technical Information ◽  
Animal Origin ◽  
Technological Properties ◽  
Recombinant Enzymes ◽  
Rennet Coagulation ◽  
Milk Coagulation

Потребность в увеличении количества и качества производимой молочной продукции стимулирует исследования, направленные на повышение эффективности переработки молочного сырья, что, в свою очередь, невозможно без изучения технологических свойств молока и функционально необходимых ингредиентов, применяемых в производстве различной молочной продукции. На основе научно-технической информации международных и российских баз данных, систематизированной в рамках изучаемой тематики, представлен обзор научных работ о коагулянтах белков молока различного происхождения, применяемых при кислотной, кислотно-сычужной и сычужной коагуляции. Установлено, что в российской и международной производственной практике востребованы коагулянты животного, микробного и рекомбинантного происхождения. Наибольшим спросом среди коагулянтов животного происхождения пользуется химозин, получаемый из желудков телят. Ферменты микробного и рекомбинантного происхождения отличаются более стабильным качеством и низкой стоимостью, но их производство, основанное на методах генной инженерии, вызывающих предубеждение у большинства потребителей, практически полностью сосредоточено за рубежом. При условии повышения лояльности потребителей ферменты рекомбинантного происхождения могут стать наиболее перспективными функциональными ингредиентами. Исследования потенциала различных протеолитических ферментов растительного происхождения выявили невысокий технологический эффект их применения. Рассмотренный материал позволяет прогнозировать перспективность научных исследований по выявлению механизмов коагуляции молока и новых эффективных и универсальных коагулянтов совокупно с селекционной практикой отбора животных, направленной на улучшение технологических свойств молочного сырья. The need to increase the quantity and quality of dairy products encourages research aimed at improving the efficiency of processing dairy raw materials, which, in turn, is impossible without studying the technological properties of milk and functionally necessary ingredients used in the production of various dairy products. On the basis of scientific and technical information from international and Russian data bases, systematized within the framework of the subject under study, a review of scientific works on milk protein coagulants of various origins used in acid, acid-rennet and rennet coagulation is presented. It is established that coagulants of animal, microbial and recombinant origin are in demand in the Russian and international production practice. The greatest demand among coagulants of animal origin is chymosin, obtained from the stomachs of calves. Microbial and recombinant enzymes are characterized by more stable quality and lower cost, but their production, based on genetic engineering methods that cause prejudice among most consumers, is almost entirely concentrated abroad. If consumer loyalty is increased, recombinant enzymes may become the most promising functional ingredients. Studies of the potential of various proteolytic enzymes of plant origin revealed a low technological effect of their use. The considered material allows us to predict the prospects of scientific research to identify the mechanisms of milk coagulation and new effective and universal coagulants together with the breeding practice of animal selection, aimed at improving the technological properties of dairy raw materials.

scholarly journals MMP20 Hemopexin Domain Mutation in Amelogenesis Imperfecta

2009 ◽  
Vol 89 (1) ◽  
pp. 46-50 ◽  
Author(s):  
S.-K. Lee ◽  
F. Seymen ◽  
H.-Y. Kang ◽  
K.-E. Lee ◽  
K. Gencay ◽  
...  
Keyword(s):  
Mutational Analysis ◽  
Proteolytic Enzymes ◽  
Dental Enamel ◽  
Amelogenesis Imperfecta ◽  
Single Amino Acid ◽  
Functional Protein ◽  
Genes Encoding ◽  
Kallikrein 4 ◽  
Normal Thickness ◽  
Hemopexin Domain

Proteolytic enzymes serve important functions during dental enamel formation, and mutations in the kallikrein 4 ( KLK4) and enamelysin ( MMP20) genes cause autosomal-recessive amelogenesis imperfecta (ARAI). So far, only 1 KLK4 and 3 MMP20 mutations have been reported in ARAI kindreds. To determine whether ARAI in a family with a hypomaturation-type enamel defect is caused by mutations in the genes encoding enamel proteolytic enzymes, we performed mutational analysis on candidate genes. Mutational and haplotype analyses revealed an ARAI-causing point mutation (c.910G>A, p.A304T) in exon 6 of MMP20 that results in a single amino acid substitution in the hemopexin domain. Western blot analysis showed decreased expression of the mutant protein, but zymogram analysis demonstrated that this mutant was a functional protein. The proband and an affected brother were homozygous for the mutation, and both unaffected parents were carriers. The enamel of newly erupted teeth had normal thickness, but was chalky white and became darker with age.

A comparative study of the porin genes encoding VDAC, a voltage-dependent anion channel protein, in Anopheles gambiae and Drosophila melanogaster

Gene ◽  
2003 ◽  
Vol 317 ◽  
pp. 111-115 ◽  
Author(s):  
Marco Sardiello ◽  
Gaetano Tripoli ◽  
Marta Oliva ◽  
Federica Santolamazza ◽  
Roberta Moschetti ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Comparative Study ◽  
Anopheles Gambiae ◽  
Anion Channel ◽  
Voltage Dependent Anion Channel ◽  
Channel Protein ◽  
Voltage Dependent ◽  
Genes Encoding

scholarly journals Comparative analysis and mutation effects of fpp2–fpp1 tandem genes encoding proteolytic extracellular enzymes of Flavobacterium psychrophilum

Microbiology ◽  
2011 ◽  
Vol 157 (4) ◽  
pp. 1196-1204 ◽  
Author(s):  
David Pérez-Pascual ◽  
Esther Gómez ◽  
Beatriz Álvarez ◽  
Jessica Méndez ◽  
Pilar Reimundo ◽  
...  
Keyword(s):  
Gene Function ◽  
Type Strain ◽  
Wild Type Strain ◽  
Proteolytic Enzymes ◽  
Function Analysis ◽  
Phenotypic Characterization ◽  
Pcr Analysis ◽  
Wild Type ◽  
Flavobacterium Psychrophilum ◽  
Genes Encoding

Flavobacterium psychrophilum is a very significant fish pathogen that secretes two biochemically characterized extracellular proteolytic enzymes, Fpp1 and Fpp2. The genes encoding these enzymes are organized as an fpp2–fpp1 tandem in the genome of strain F. psychrophilum THC02/90. Analysis of the corresponding encoded proteins showed that they belong to two different protease families. For gene function analysis, new genetic tools were developed in F. psychrophilum by constructing stable isogenic fpp1 and fpp2 mutants via single-crossover homologous recombination. RT-PCR analysis of wild-type and mutant strains suggested that both genes are transcribed as a single mRNA from the promoter located upstream of the fpp2 gene. Phenotypic characterization of the fpp2 mutant showed lack of caseinolytic activity and higher colony spreading compared with the wild-type strain. Both characteristics were recovered in the complemented strain. One objective of this work was to assess the contribution to virulence of these proteolytic enzymes. LD50 experiments using the wild-type strain and mutants showed no significant differences in virulence in a rainbow trout challenge model, suggesting instead a possible nutritional role. The gene disruption procedure developed in this work, together with the knowledge of the complete genome sequence of F. psychrophilum, open new perspectives for the study of gene function in this bacterium.

scholarly journals Enzymatic Detachment of Therapeutic Mesenchymal Stromal Cells Grown on Glass Carriers in a Bioreactor

2013 ◽  
Vol 7 (1) ◽  
pp. 147-158 ◽  
Author(s):  
Denise Salzig ◽  
Alexandra Schmiermund ◽  
Pablo P. Grace ◽  
Christiane Elseberg ◽  
Christian Weber ◽  
...  
Keyword(s):  
Mesenchymal Stromal Cells ◽  
Dynamic Systems ◽  
Stromal Cells ◽  
Proteolytic Enzymes ◽  
Cell Damage ◽  
Animal Origin ◽  
Cell Detachment ◽  
Adherent Cells ◽  
Experimental Conditions ◽  
Cell Harvest

Cell therapies require the in vitro expansion of adherent cells such as mesenchymal stromal cells (hMSCs) in bioreactor systems or other culture environments, followed by cell harvest. As hMSCs are strictly adherent cells, cell harvest requires cell detachment. The use of hMSCs for cell therapy requires GMP production in accordance with the guidelines for advanced therapeutic medical products. Therefore, several GMP-conform available proteolytic enzymes were investigated for their ability to promote hMSC detachment. An allogeneic hMSC cell line (hMSC-TERT) that is used in clinical trials in the form of alginate cell capsules was chosen as a model. This study investigated the influence of several factors on the outcome of proteolytic hMSC-TERT detachment. Therefore, hMSC-TERT detachment was analyzed in different cultivation systems (static, dynamic) and in combination with further cell processing including encapsulation. Only two of the commercially available enzymes (AccutaseTM, TrypZeanTM) that fulfill all process requirements (commercial availability, cost, GMP conditions during manufacturing and non-animal origin) are found to be generally suitable for detaching hMSC-TERT. Combining cell detachment with encapsulation demonstrated a high impact of the experimental set up on cell damage. It was preferable to reduce the temperature during detachment and limit the detachment time to a maximum of 20 minutes. Cell detachment in static systems was not comparable with detachment in dynamic systems. Detachment yields in dynamic systems were lower and cell damage was higher for the same experimental conditions. Finally, only TrypZeanTM seemed to be suitable for the detachment of hMSC-TERT from dynamic reactor systems.

scholarly journals A locus on human chromosome 20 contains several genes expressing protease inhibitor domains with homology to whey acidic protein

2002 ◽  
Vol 368 (1) ◽  
pp. 233-242 ◽  
Author(s):  
Adam CLAUSS ◽  
Hans LILJA ◽  
Åke LUNDWALL
Keyword(s):  
Human Chromosome ◽  
Protease Inhibitor ◽  
Proteolytic Enzymes ◽  
Whey Acidic Protein ◽  
Acidic Protein ◽  
Promoter Regions ◽  
Transcript Levels ◽  
Genes Encoding ◽  
Characteristic Features ◽  
Rapid Amplification

A locus containing 14 genes, encoding protein domains that have homology with whey acidic protein (WAP), has been identified in a region of 678kb on human chromosome 20q12-13.1. Among them are genes of the known or postulated protease inhibitors elafin, secretory leucocyte protease inhibitor, human epididymis gene product 4, eppin, and huWAP2. Nucleotide sequences of full-length transcripts were obtained from cDNA fragments generated by rapid amplification of cDNA ends. Characteristic features of the genes are that the upstream promoter regions are devoid of TATA-boxes and that the coding nucleotides are divided into distinct exons for the signal peptide and for each WAP domain. In most cases, there is also a separate exon encompassing a few terminal codons and the 3′ untranslated nucleotides. There are also examples of mixed type inhibitors, that encode inhibitor domains of both WAP and Kunitz types. Several of the genes appear to be expressed ubiquitously, but, in most cases, the highest transcript levels are found in epididymis followed by testis and trachea. Some of the genes also display high transcript levels in neural tissues. Potential biological roles of protein products could be in host defence against invading micro-organisms or in the regulation of endogenous proteolytic enzymes, of which those originating from the kallikrein gene locus on chromosome 19 are of particular interest.

Genetic characterization of staphopain genes in Staphylococcus aureus

2004 ◽  
Vol 385 (11) ◽  
pp. 1059-1067 ◽  
Author(s):  
Ewa Golonka ◽  
Renata Filipek ◽  
Artur Sabat ◽  
Anna Sinczak ◽  
Jan Potempa
Keyword(s):  
Staphylococcus Aureus ◽  
Bacterial Infections ◽  
Genomic Sequence ◽  
Sequence Data ◽  
Proteolytic Enzymes ◽  
Disease Process ◽  
Pcr Amplification ◽  
Single Copy ◽  
Pcr Rflp ◽  
Genes Encoding

AbstractStaphylococcus aureus, a leading cause of bacterial infections in humans, is endowed with a wealth of virulence factors that contribute to the disease process. Several extracellular proteolytic enzymes, including cysteine proteinases referred to as the staphopains (staphopain A, encoded by thescpAgene, and staphopain B, encoded bysspB), have proposed roles for staphylococcal virulence. Here we present data regarding the distribution, copy number and genetic variability of the genes encoding the staphopains in a large number ofS. aureusstrains. The polymorphism of thescpAandsspBgenes in three laboratory strains and 126 clinical isolates was analyzed by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP). Both genes were detected in all isolates by PCR amplification and, based on the PCR-RFLP patterns, classified as four types forscpAand six types forsspB. Those with the most divergent patterns were subjected to DNA sequencing and compared with genomic sequence data for the seven available strains ofS. aureus. Southern blot analysis of thescpAandsspBsequences indicates that they are strongly conserved as single-copy genes in the genome of eachS. aureusstrain investigated. Taken together, these data suggest that the staphopains have important housekeeping and/or virulence functions, and therefore may constitute an interesting target for the development of therapeutic inhibitors for the treatment of staphylococcal diseases.

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