scholarly journals Functional metagenomics using Pseudomonas putida expands the known diversity of polyhydroxyalkanoate synthases and enables the production of novel polyhydroxyalkanoate copolymers

2016 ◽  
Author(s):  
Jiujun Cheng ◽  
Trevor Charles

Bacterially produced biodegradable polyhydroxyalkanoates with versatile properties can be achieved using different PHA synthase enzymes. This work aims to expand the diversity of known PHA synthases via functional metagenomics, and demonstrates the use of these novel enzymes in PHA production. Complementation of a PHA synthesis deficient Pseudomonas putida strain with a soil metagenomic cosmid library retrieved 27 clones expressing either Class I, Class II or unclassified PHA synthases, and many did not have close sequence matches to known PHA synthases. The composition of PHA produced by these clones was dependent on both the supplied growth substrates and the nature of the PHA synthase, with various combinations of SCL- and MCL-PHA. These data demonstrate the ability to isolate diverse genes for PHA synthesis by functional metagenomics, and their use for the production of a variety of PHA polymer and copolymer mixtures.

2019 ◽  
Vol 20 (5) ◽  
Author(s):  
WATUMESA A TAN ◽  
IRA WIJAYA ◽  
TRESNAWATI PURWADARIA

Abstract. Tan WA, Wijaya I, Purwadaria T. 2019. Bioprospecting of polyhydroxyalkanoates-producing bacteria from Indonesian marine environment. Biodiversitas 20: 1309-1315. Polyhydroxyalkanoates (PHA) are potential alternates to conventional synthetic plastics. PHA production in bacteria involves PHA synthase gene encoded by phaC. In this study, we isolated PHA-producing bacteria from the coastline and 1 mile from the coastline of three beaches in Indonesia. Further phaC detection and characterization of PHA production were conducted. The isolates were subjected to phylogenetic analysis based on 16S rDNA. Red Nile staining on minimal agar revealed that twenty-three isolates showed orange fluorescent, which indicated that they accumulated PHA in their cells. PCR detection showed the presence of PHA synthase class I-encoding gene phaC in twelve isolates. One representative amplicon was sequenced to verify its identity, in which it shared 86% similarity with the PHA synthase class I-encoding gene from an uncultured bacterium. Interestingly, the production of PHA in isolate ST.PA.75, which was closely related to Vibrio sp., was 2.1-fold higher than that in the Ralstonia eutropha JMP134 control. Three isolates showed similarity with bacterial genera and/or species for which PHA producing phenotypes had never been described before TP.SWC.20, which was closely related to Microbacterium arborescens, as well as TP.SWC.33 and TP.SWC.85, which were similar to Psychrobacter spp. Phylogenetic analysis showed that the PHA producing isolates were clustered into three phyla: γ-Proteobacteria, Actinomycetes, and Bacilli. A majority of the isolates (75%) were related to γ-Proteobacteria. In this study, we uncovered diverse novel promising strains for use in the production of PHA as a more environmentally-friendly alternative to hydrocarbon-based plastics.


Author(s):  
T. A. Stewart ◽  
D. Liggitt ◽  
S. Pitts ◽  
L. Martin ◽  
M. Siegel ◽  
...  

Insulin-dependant (Type I) diabetes mellitus (IDDM) is a metabolic disorder resulting from the lack of endogenous insulin secretion. The disease is thought to result from the autoimmune mediated destruction of the insulin producing ß cells within the islets of Langerhans. The disease process is probably triggered by environmental agents, e.g. virus or chemical toxins on a background of genetic susceptibility associated with particular alleles within the major histocompatiblity complex (MHC). The relation between IDDM and the MHC locus has been reinforced by the demonstration of both class I and class II MHC proteins on the surface of ß cells from newly diagnosed patients as well as mounting evidence that IDDM has an autoimmune pathogenesis. In 1984, a series of observations were used to advance a hypothesis, in which it was suggested that aberrant expression of class II MHC molecules, perhaps induced by gamma-interferon (IFN γ) could present self antigens and initiate an autoimmune disease. We have tested some aspects of this model and demonstrated that expression of IFN γ by pancreatic ß cells can initiate an inflammatory destruction of both the islets and pancreas and does lead to IDDM.


1991 ◽  
Vol 17 (1) ◽  
pp. 53-62
Author(s):  
Irene Hughson

Summary This paper examines the horse carvings to be found on Class I and Class II Pictish sculptured stones and considers their reliability as evidence of the sort of horses and ponies that would have existed in the Early Historic Period. An attempt is made to show that the availability in Britain of good sized, high quality riding horses during that period is not inconsistent with what is known of the development and distribution of different types of horses in pre-hislory. The importance of horses and ponies in Early Historic societies is stressed and inferences drawn about the agricultural economy that could support horses and the skilled specialists required to look after them.


2020 ◽  
Vol 8 (3) ◽  
pp. 144-156
Author(s):  
Şule KARATAŞ ◽  
Fatma SAVRAN OĞUZ

Introduction: Peptides obtained by processing intracellular and extracellular antigens are presented to T cells to stimulate the immune response. This presentation is made by peptide receptors called major histocompatibility complex (MHC) molecules. The regulation mechanisms of MHC molecules, which have similar roles in the immune response, especially at the gene level, have significant differences according to their class. Objective: Class I and class II MHC molecules encoded by MHC genes on the short arm of the sixth chromosome are peptide receptors that stimulate T cell response. These peptides, which will enable the recognition of the antigen from which they originate, are loaded into MHC molecules and presented to T cells. Although the principles of loading and delivering peptides are similar for both molecules, the peptide sources and peptide loading mechanisms are different. In addition, class I molecules are expressed in all nucleated cells while class II molecules are expressed only in Antigen Presentation Cells (APC). These differences; It shows that MHC class I is not expressed by exactly the same transcriptional mechanisms as MHC class II. In our article, we aimed to compare the gene expressions of both classes and reveal their similarities and differences. Discussion and Conclusion: A better understanding of the transcriptional mechanisms of MHC molecules will reveal the role of these molecules in diseases more clearly. In our review, we discussed MHC gene regulation mechanisms with presence of existing informations, which is specific to the MHC class, for contribute to future research. Keywords: MHC class I, MHC class II, MHC gene regulation, promoter, SXY module, transcription


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