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Glycobiology Sep 2019The development of therapeutic proteins for the treatment of numerous diseases is one of the fastest growing areas of biotechnology. Therapeutic efficacy and serum...
The development of therapeutic proteins for the treatment of numerous diseases is one of the fastest growing areas of biotechnology. Therapeutic efficacy and serum half-life are particularly important, and these properties rely heavily on the glycosylation state of the protein. Expression systems to produce authentically fully glycosylated therapeutic proteins with appropriate terminal sialic acids are not yet perfected. The in vitro modification of therapeutic proteins by recombinant sialyltransferases offers a promising and elegant strategy to overcome this problem. Thus, the detailed expression and characterization of sialyltransferases for completion of the glycan chains is of great interest to the community. We identified a novel α2,6-sialyltransferase from Helicobacter cetorum and compared it to the human ST6Gal1 and a Photobacterium sp. sialyltransferase using glycoprotein substrates in a 96-well microtiter-plate-based assay. We demonstrated that the recombinant α2,6-sialyltransferase from H. cetorum is an excellent catalyst for modification of N-linked glycans of different therapeutic proteins.
Topics: Antigens, CD; Cloning, Molecular; Glycoproteins; Glycosylation; Helicobacter; Humans; Photobacterium; Polysaccharides; Protein Processing, Post-Translational; Sialic Acids; Sialyltransferases; beta-D-Galactoside alpha 2-6-Sialyltransferase
PubMed: 31281932
DOI: 10.1093/glycob/cwz050 -
International Journal of Molecular... Aug 2022The genus is known for its ecophysiological versatility encompassing free-living, symbiotic, and pathogenic lifestyles. sp. CCB-ST2H9 was isolated from estuarine...
The genus is known for its ecophysiological versatility encompassing free-living, symbiotic, and pathogenic lifestyles. sp. CCB-ST2H9 was isolated from estuarine sediment collected at Matang Mangrove, Malaysia. In this study, the genome of CCB-ST2H9 was sequenced, and the pan-genome of 37 strains was analysed. Phylogeny based on core genes showed that CCB-ST2H9 clustered with , forming a distinct clade with , , and . The core genome of was conserved in housekeeping functions, while the flexible genome was well represented by environmental genes related to energy production and carbohydrate metabolism. Genomic metrics including 16S rRNA sequence similarity, average nucleotide identity, and digital DNA-DNA hybridization values were below the cut-off for species delineation, implying that CCB-ST2H9 potentially represents a new species. Genome mining revealed that biosynthetic gene clusters (BGCs) involved in producing antimicrobial compounds such as holomycin in CCB-ST2H9 could contribute to the antagonistic potential. Furthermore, the EtOAc extract from the culture broth of CCB-ST2H9 exhibited antagonistic activity against spp. Intriguingly, clustering based on BGCs profiles grouped , , , , and CCB-ST2H9 together in the heatmap by the presence of a large number of BGCs. These BGCs-rich strains represent great potential for bioactive secondary metabolites production and sources for novel compounds.
Topics: Bacterial Typing Techniques; Base Composition; DNA, Bacterial; Fatty Acids; Genomics; Multigene Family; Photobacterium; Phylogeny; RNA, Ribosomal, 16S; Sequence Analysis, DNA
PubMed: 36077108
DOI: 10.3390/ijms23179712 -
International Journal of Molecular... Jul 2022The evaluation of temperature effects on the structure and function of enzymes is necessary to understand the mechanisms underlying their adaptation to a constantly...
The evaluation of temperature effects on the structure and function of enzymes is necessary to understand the mechanisms underlying their adaptation to a constantly changing environment. In the current study, we investigated the influence of temperature variation on the activity, structural dynamics, thermal inactivation and denaturation of and luciferases belonging to different subfamilies, as well as the role of sucrose in maintaining the enzymes functioning and stability. We used the stopped-flow technique, differential scanning calorimetry and molecular dynamics to study the activity, inactivation rate, denaturation and structural features of the enzymes under various temperatures. It was found that luciferase resembles the properties of cold-adapted enzymes with high activity in a narrow temperature range and slightly lower thermal stability than luciferase, which is less active, but more thermostable. Differences in activity at the studied temperatures can be associated with the peculiarities of the mobile loop conformational changes. The presence of sucrose does not provide an advantage in activity but increases the stability of the enzymes. Differential scanning calorimetry experiments showed that luciferases probably follow different denaturation schemes.
Topics: Luciferases; Luciferases, Bacterial; Structure-Activity Relationship; Sucrose; Temperature
PubMed: 35897698
DOI: 10.3390/ijms23158119 -
Frontiers in Oncology 2022Commensal microbiota have been proven to colonize the mammary gland, but whether their composition is altered in patients with breast cancer (BC) remains elusive. This...
BACKGROUND
Commensal microbiota have been proven to colonize the mammary gland, but whether their composition is altered in patients with breast cancer (BC) remains elusive. This study intends to explore the breast microbiome differences between benign and malignant diseases and to investigate the impact of neoadjuvant chemotherapy (NAC) on the breast microbiota in patients with BC.
METHODS
Breast normal adipose tissues (NATs) were collected from 79 patients with BC and 15 controls between July 2019 and November 2021. The BC group consisted of 29 patients who had received NAC and 50 who were non-NAC patients. Participants diagnosed with benign breast disease were recruited as controls. 16S rRNA gene sequencing was used to analyze the bacterial diversity of NATs.
RESULTS
The community structure of the NAT microbiome was significantly different between the BC and control groups. decreased (47.40% versus 39.74%), whereas increased (15.71% versus 25.33%) in patients with BC when compared with that in control tissues. Nine genera were enriched in BC NATs, and four genera levels increased in the control group. The associations between differential bacterial genera and breast tumor grade were calculated by Spearman's correlation. The results showed that tumor grade was positively associated with the relative abundance of and negatively related to , , , and . Moreover, menopause was associated with the microbiota composition change of non-NAC BC patients and related to the significant reduction in the abundance level of , , and . In addition, NAC was related to the beta diversity of patients with BC and associated with the decrease of and in postmenopausal patients. Of note, Tax4Fun functional prediction analysis revealed that the metabolic state was more exuberant in the BC group with upregulating of multiple metabolism-related pathways.
CONCLUSION
Our results offer new insight into the relationship between NAC and breast microbiota and help to better characterize the breast microbial dysbiosis that occurs in patients with BC. Further epidemiological studies with larger sample size and well-designed animal experiments are required to elucidate the role of breast microbiota in the therapeutic outcome of BC.
PubMed: 36172155
DOI: 10.3389/fonc.2022.926920 -
FEMS Microbiology Ecology Dec 2022The occurrence of pathogenic bacteria has emerged as a plausible key component of summer mortalities in mussels. In the current research, four bacterial isolates...
The occurrence of pathogenic bacteria has emerged as a plausible key component of summer mortalities in mussels. In the current research, four bacterial isolates retrieved from moribund Greenshell࣪ mussels, Perna canaliculus, from a previous summer mortality event, were tentatively identified as Vibrio and Photobacterium species using morpho-biochemical characterization and MALDI-TOF MS and confirmed as V. celticus, P. swingsii, P. rosenbergii, and P. proteolyticum using whole genome sequencing. These isolates were utilized in a laboratory challenge where mussels were injected with cell concentrations ranging from 105 to 109 CFU/mussel. Of the investigated isolates, P. swingsii induced the highest mortality. Additionally, results from quantitative polymerase chain reaction analysis, focusing on known virulence genes were detected in all isolates grown under laboratory conditions. Photobacterium rosenbergii and P. swingsii showed the highest expression levels of these virulence determinants. These results indicate that Photobacterium spp. could be a significant pathogen of P. canaliculus, with possible importance during summer mortality events. By implementing screening methods to detect and monitor Photobacterium concentrations in farmed mussel populations, a better understanding of the host-pathogen relationship can be obtained, aiding the development of a resilient industry in a changing environment.
Topics: Animals; Perna; Vibrio; Seasons; Virulence Factors; Seafood
PubMed: 36449667
DOI: 10.1093/femsec/fiac140 -
Microbiological Research Mar 2020Photobacterium species are widely distributed in the marine environment. The overall metabolism of this genus remains largely unknown. In order to improve our knowledge...
Photobacterium species are widely distributed in the marine environment. The overall metabolism of this genus remains largely unknown. In order to improve our knowledge on this bacterium, the relationship between the genome and phenome of the Photobacterium isolate was analyzed. The cream colored, Gram-negative, rod-shaped and motile bacterial strain, J15, was isolated from marine water of Tanjung Pelepas, Johor, Malaysia. The 5,684,538 bp genome of strain J15 comprised 3 contigs (2 chromosomes and 1 plasmid) with G + C content of 46.39 % and contained 4924 protein-coding genes including 180 tRNAs and 40 rRNAs. The phenotypic microarray (PM) as analyzed using BIOLOG showed the utilization of; i) 93 of the 190 carbon sources tested, where 61 compounds were used efficiently; ii) 41 of the 95 nitrogen sources tested, where 22 compounds were used efficiently; and iii) 3 of the 94 phosphorous and sulphur sources tested. Furthermore, high tolerance to osmotic stress, basic pH and toxic compounds as well as resistance to antibiotics of strain J15 were determined by BIOLOG PM. The ANI and kSNP analyses revealed that strain J15 to be the same species with Photobacterium marinum AK15 with ANI value of 96.93 % and bootstrapping value of 100 in kSNP. Based on the ANI and kSNP analyses, strain J15 was identified as P. marinum J15.
Topics: Aquatic Organisms; Base Composition; DNA, Bacterial; Genome, Bacterial; Genomics; Malaysia; Phenomics; Photobacterium; Phylogeny; Seawater; Sequence Analysis, DNA
PubMed: 31945517
DOI: 10.1016/j.micres.2020.126410 -
Molecules (Basel, Switzerland) Jul 2022This study aimed to evaluate the antibacterial activity in vitro of and extracts against potentially pathogenic bacteria for Pacific white shrimp. Furthermore, the...
This study aimed to evaluate the antibacterial activity in vitro of and extracts against potentially pathogenic bacteria for Pacific white shrimp. Furthermore, the extracts with higher inhibitory activity were analyzed to identify compounds responsible for bacterial inhibition and evaluate their effect on motility and biofilm formation. and extracts were prepared using methanol, acetone, and hexane by ultrasound. The minimum inhibitory concentration (MIC) of the extracts was determined against and . The polyphenol profile of those extracts showing the highest bacterial inhibition were determined. Besides, the bacterial swimming and swarming motility and biofilm formation were determined. The highest inhibitory activity against the four pathogens was found with the acetonic extract of leaf (MIC of 50 mg/mL for spp. and 25 mg/mL for spp.) and the methanol extract of flower (MIC of 50 mg/mL for all pathogens tested). Both extracts affected the swarming and swimming motility and the biofilm formation of the tested bacteria. The main phenolic compounds related to bacteria inhibition were naringin, vanillic acid, and rosmarinic acid, whilst hesperidin, kaempferol pentosyl-rutinoside, and rhamnetin were related to bacteria inhibition.
Topics: Animals; Anti-Bacterial Agents; Methanol; Microbial Sensitivity Tests; Penaeidae; Plant Extracts; Vibrio parahaemolyticus
PubMed: 35889267
DOI: 10.3390/molecules27144397 -
Frontiers in Immunology 2021Cobia () is a marine fish of high economic value that grows at a fast rate. However, intensive fish farming has led to disease outbreaks in cobia cultures, which is...
Cobia () is a marine fish of high economic value that grows at a fast rate. However, intensive fish farming has led to disease outbreaks in cobia cultures, which is highly costly to the industry. The impact of infectious diseases on cobia production has led to the inappropriate and increased use of chemicals and antibiotics, which negatively affects the environment and human health and promotes the spread of drug-resistant pathogens. Hence, prophylactic measurements, such as the use of immunomodulators, are required to improve the health of cultured animals against pathogens. In this study, we examined the effects of water extract (SSWE) in cobia and . We found that treatment with SSWE could significantly increase the expression of cytokines (e.g., , , , , and ) and chemokines (e.g., ) in primary cultured head kidney leukocytes. Intraperitoneal injection of SSWE (20 μg/g body weight) promoted higher expression of , , , , chemokines (e.g., ), and antibodies (e.g., ) in head kidney and spleen tissues of the fish compared with other dose levels. Additionally, we describe for the second time (only after India) of the isolation of subsp. () from a deadly epizootic in cage-farmed cobia. An intraperitoneal inoculation of SSWE before challenge showed that SSWE treatment could delay the onset of mortality of cobia. Finally, fish that received SSWE intraperitoneally before infection with exhibited elevated expression of Th1-type cytokines, namely, , , , and . At the same time, the expression of Th2-related factors (such as in the head kidney, and and in the spleen) were lower for the fish that received SSWE instead of PBS before the challenge. The results indicate that SSWE treatment facilitates the induction of Th1-type cytokines in cobia to fight against infection and has the potential to be used as an immunostimulant and vaccine adjuvant for fish.
Topics: Animals; Biological Products; Cytokines; Fish Diseases; Host-Pathogen Interactions; Immunity, Innate; Inflammation Mediators; Mortality; Photobacterium; Rhodophyta; Th1 Cells; Transcriptome
PubMed: 35140710
DOI: 10.3389/fimmu.2021.801501 -
Applied and Environmental Microbiology Jul 2023Coral reefs are among the most biodiverse ecosystems, providing habitats for various organisms. Studies on coral bleaching have been increasing recently, but little is...
Coral reefs are among the most biodiverse ecosystems, providing habitats for various organisms. Studies on coral bleaching have been increasing recently, but little is known about the distribution and community assembly of coral pathogenic bacteria (e.g., several species). We elucidated the distribution pattern and interaction relationships of total bacteria and spp. in sediments from the Xisha Islands, which are characterized by their high coverage and diversity of coral resources. spp. showed significantly higher relative abundance values in the Xisha Islands (1.00 × 10 copies/g) than in other areas (approximately 1 × 10 to 9.04 × 10 copies/g), indicating that the coral bleaching event of 2020 may have promoted the bloom of vibrios. A spatial shift in community composition was observed between the northern (Photobacterium rosenbergii and Vibrio ponticus) and southern (Vibrio ishigakensis and Vibrio natriegens) sites, accompanied by a clear distance-decay pattern. The spatial distance and coral species (e.g., and ) had much greater correlations with the community than did environmental factors. However, complex mechanisms may exist in the community assembly of spp. due to the large proportion of unexplained variation. Stochastic processes may play an important role, as shown by the neutral model. Vibrio harveyi had the highest relative abundance (77.56%) and niche breadth, compared to other species, and it was negatively correlated with Acroporidae, likely reflecting its strong competitive ability and adverse effects on specific corals. Our study provides insights into the bloom and underlying assembly mechanisms of sedimentary vibrios in the Xisha Islands, thereby contributing to identify the potential indicator of coral bleaching and provide inspiration for the environmental management of coral reef areas. Coral reefs exert important roles in maintaining the sustainability of marine ecosystems but decline worldwide due to various drivers, especially pathogenic microorganisms. Here, we investigated the distribution pattern and interactions of total bacteria and spp. in the sediments from Xisha Islands during the coral bleaching event of 2020. Our results showed that the abundances of (1.00 × 10 copies/g) were high across the whole sites, indicating the bloom of sedimentary spp. Coral pathogenic species were abundant in the sediments, likely reflecting adverse effects on several kinds of corals. The compositions of the spp. were separated by geographical location, which was mainly attributable to the spatial distance and coral species. Overall, this work contributes by providing evidence for the outbreak of coral pathogenic vibrios. The pathogenic mechanism of the dominant species (especially V. harveyi) should be comprehensively considered by laboratory infection experiments in the future.
Topics: Animals; Ecosystem; Coral Bleaching; Islands; Coral Reefs; Anthozoa; Vibrio
PubMed: 37314342
DOI: 10.1128/aem.00543-23 -
Animal Nutrition (Zhongguo Xu Mu Shou... Jun 2022The utilization efficiency of soy protein is affected by its 2 anti-nutritional substances-the antigens β-conglycinin and glycinin. This study investigated their...
The utilization efficiency of soy protein is affected by its 2 anti-nutritional substances-the antigens β-conglycinin and glycinin. This study investigated their effects on the growth performance, intestinal immune defense, and microbiome in juvenile pearl gentian groupers (♀ × ♂). Three isonitrogenous and isolipidic diets were formulated containing fishmeal supplemented with 70 g/kg β-conglycinin or 100 g/kg glycinin, or no supplementation (control). Each experimental diet was fed to quadruplicate groups with 30 fish in each tank for 8 weeks. Dietary inclusion of either β-conglycinin or glycinin significantly reduced weight gain and specific growth rates, and cell proliferation of the distal intestine. Histological evaluation of the intestine tract revealed the inflammation signs, characterized by reducing of plica height and width as well as the number of the goblet cells, and widening of the lamina propria. The group fed the β-conglycinin diet had reduced lysozyme activity, contents of immunoglobulin M and complements 3 and 4. Increased activities of caspase-3 and -9 were observed in the group fed the β-conglycinin diet compared to the other 2 groups. In the intestinal microbiota, the relative abundances of the potentially pathogenic genera and were significantly higher in the glycinin group than those in others. Therefore, the existence of soybean antigens (β-conglycinin or glycinin) could damage the structural integrity of the intestine, reduce immune defense, reshape the intestinal microbiome and, ultimately, impair growth in fish.
PubMed: 35600546
DOI: 10.1016/j.aninu.2021.11.001