scholarly journals Pectin lyase production by recombinant Penicillium griseoroseum strain 105

2010 ◽  
Vol 56 (10) ◽  
pp. 831-837 ◽  
Author(s):  
Patrícia Gomes Cardoso ◽  
Janaina Aparecida Teixeira ◽  
Marisa Vieira de Queiroz ◽  
Elza Fernandes de Araújo

Recombinant Penicillium griseoroseum strain 105 overproduces an extracellular pectin lyase (PL) under the transcriptional control of the strong gpdA promoter of Aspergillus nidulans . Our aim was to evaluate PL production by recombinant P. griseoroseum strain 105 in submerged fermentation system bioreactors BioFloIII and BioFloIV using 2 or 10 L working volumes under different growth conditions and to analyze the production of cellulase, polygalacturonase, pectin methylesterase, and protease. PL overproduction by recombinant P. griseoroseum strain 105 was 112 times higher than that of P. griseoroseum PG63 grown in sugarcane juice. Cellulases and proteases were not detected in the culture filtrate, and evaluation for extracellular proteins in the culture medium by SDS–PAGE showed the presence of a 36 kDa predominant band, similar to the molecular mass estimated from the nucleotide sequence of plg1 gene for PL of P. griseoroseum strain 105. This recombinant strain provides the advantage of PL production, which predominates over other extracellular proteins usually present in most commercial pectinase preparations, using sugarcane juice as a substrate of low cost.

Author(s):  
Mostafa Mabrouk ◽  
Tarek A. Elkhooly ◽  
Shaimaa K. Amer

Abstract Background Bio-nanotechnology is considered as one of the low-cost approaches that have been utilized in production of nanomaterials. The current research aimed at investigating the influence of different types of Actinomycete strains on the final properties of silver nanoparticles (AgNPs) such as size, shape, polydispersity, and antibacterial properties. For this purpose, the following techniques were employed UV spectrophotometer, SDS-PAGE electrophoresis, TEM, FTIR, antibacterial agar diffusion test, and Zetasizer. Results It was found that among 34 Streptomyces isolates collected from the soil, Streptomyces spiralis and Streptomyces rochei were able to reduce silver nitrate into sliver nanoparticles. The diversity and molecular weights of extracellular proteins secreted by these stains were different as proved by SDS-PAGE technique. This consequently resulted in differences in polydispersity of AgNPs which indicate that the sizes of AgNPs were highly dependent on the amount, molecular sizes, and diversity of extracellular matrix proteins of the microorganism. Conclusion This article might give an insight about the importance of molecular sizes of biomacromolecules such as proteins on the physical properties of biogenic synthesized nanoparticles.


Author(s):  
K.M. Hones ◽  
P. Sheldon ◽  
B.G. Yacobi ◽  
A. Mason

There is increasing interest in growing epitaxial GaAs on Si substrates. Such a device structure would allow low-cost substrates to be used for high-efficiency cascade- junction solar cells. However, high-defect densities may result from the large lattice mismatch (∼4%) between the GaAs epilayer and the silicon substrate. These defects can act as nonradiative recombination centers that can degrade the optical and electrical properties of the epitaxially grown GaAs. For this reason, it is important to optimize epilayer growth conditions in order to minimize resulting dislocation densities. The purpose of this paper is to provide an indication of the quality of the epitaxially grown GaAs layers by using transmission electron microscopy (TEM) to examine dislocation type and density as a function of various growth conditions. In this study an intermediate Ge layer was used to avoid nucleation difficulties observed for GaAs growth directly on Si substrates. GaAs/Ge epilayers were grown by molecular beam epitaxy (MBE) on Si substrates in a manner similar to that described previously.


2021 ◽  
pp. 088391152199784
Author(s):  
Nipun Jain ◽  
Shashi Singh

Development of an artificial tissue by tissue engineering is witnessed to be one of the long lasting clarified solutions for the damaged tissue function restoration. To accomplish this, a scaffold is designed as a cell carrier in which the extracellular matrix (ECM) performs a prominent task of controlling the inoculated cell’s destiny. ECM composition, topography and mechanical properties lead to different types of interactions between cells and ECM components that trigger an assortment of cellular reactions via diverse sensing mechanisms and downstream signaling pathways. The polysaccharides in the form of proteoglycans and glycoproteins yield better outcomes when included in the designed matrices. Glycosaminoglycan (GAG) chains present on proteoglycans show a wide range of operations such as sequestering of critical effector morphogens which encourage proficient nutrient contribution toward the growing stem cells for their development and endurance. In this review we discuss how the glycosylation aspects are of considerable importance in everyday housekeeping functions of a cell especially when placed in a controlled environment under ideal growth conditions. Hydrogels made from these GAG chains have been used extensively as a resorbable material that mimics the natural ECM functions for an efficient control over cell attachment, permeability, viability, proliferation, and differentiation processes. Also the incorporation of non-mammalian polysaccharides can elicit specific receptor responses which authorize the creation of numerous vigorous frameworks while prolonging the low cost and immunogenicity of the substance.


2021 ◽  
Author(s):  
Nick Fackler ◽  
James Heffernan ◽  
Alex Juminaga ◽  
Damien Doser ◽  
Shilpa Nagaraju ◽  
...  

Abstract Gas fermentation by Clostridium autoethanogenum is a commercial process for the sustainable biomanufacturing of fuels and valuable chemicals using abundant, low cost C1 feedstocks (CO and CO2) from sources such as inedible biomass, unsorted and non-recyclable municipal solid waste, and industrial emissions. Efforts towards pathway engineering and elucidation of gene function in this microbe have been limited by a lack of genetic tools to control gene expression and arduous genome engineering methods. To increase the pace of progress, here we developed an inducible CRISPR interference (CRISPRi) system for C. autoethanogenum and applied that system towards transcriptional repression of genes with ostensibly crucial functions in metabolism.


1999 ◽  
Vol 76 (2) ◽  
pp. 129-142 ◽  
Author(s):  
Maria Cristina Baracat-Pereira ◽  
Jorge Luiz Cavalcante Coelho ◽  
Rosana Cristina Minussi ◽  
Virgínia Maria Chaves-Alves ◽  
Rogelio Lopes Brandio ◽  
...  

2013 ◽  
Vol 85 (4) ◽  
pp. 1427-1438 ◽  
Author(s):  
MATHIAS A. CHIA ◽  
ANA T. LOMBARDI ◽  
MARIA DA GRACA G. MELAO

The need for clean and low-cost algae production demands for investigations on algal physiological response under different growth conditions. In this research, we investigated the growth, biomass production and biochemical composition of Chlorella vulgaris using semi-continuous cultures employing three growth media (LC Oligo, Chu 10 and WC media). The highest cell density was obtained in LC Oligo, while the lowest in Chu medium. Chlorophyll a, carbohydrate and protein concentrations and yield were highest in Chu and LC Oligo media. Lipid class analysis showed that hydrocarbons (HC), sterol esthers (SE), free fatty acids (FFA), aliphatic alcohols (ALC), acetone mobile polar lipids (AMPL) and phospholipids (PL) concentrations and yields were highest in the Chu medium. Triglyceride (TAG) and sterol (ST) concentrations were highest in the LC Oligo medium. The results suggested that for cost effective cultivation, LC Oligo medium is the best choice among those studied, as it saved the cost of buying vitamins and EDTA associated with the other growth media, while at the same time resulted in the best growth performance and biomass production.


Plant Methods ◽  
2019 ◽  
Vol 15 (1) ◽  
Author(s):  
Dominik Schneider ◽  
Laura S. Lopez ◽  
Meng Li ◽  
Joseph D. Crawford ◽  
Helmut Kirchhoff ◽  
...  

Abstract Background Over the last years, several plant science labs have started to employ fluctuating growth light conditions to simulate natural light regimes more closely. Many plant mutants reveal quantifiable effects under fluctuating light despite being indistinguishable from wild-type plants under standard constant light. Moreover, many subtle plant phenotypes become intensified and thus can be studied in more detail. This observation has caused a paradigm shift within the photosynthesis research community and an increasing number of scientists are interested in using fluctuating light growth conditions. However, high installation costs for commercial controllable LED setups as well as costly phenotyping equipment can make it hard for small academic groups to compete in this emerging field. Results We show a simple do-it-yourself approach to enable fluctuating light growth experiments. Our results using previously published fluctuating light sensitive mutants, stn7 and pgr5, confirm that our low-cost setup yields similar results as top-prized commercial growth regimes. Moreover, we show how we increased the throughput of our Walz IMAGING-PAM, also found in many other departments around the world. We have designed a Python and R-based open source toolkit that allows for semi-automated sample segmentation and data analysis thereby reducing the processing bottleneck of large experimental datasets. We provide detailed instructions on how to build and functionally test each setup. Conclusions With material costs well below USD$1000, it is possible to setup a fluctuating light rack including a constant light control shelf for comparison. This allows more scientists to perform experiments closer to natural light conditions and contribute to an emerging research field. A small addition to the IMAGING-PAM hardware not only increases sample throughput but also enables larger-scale plant phenotyping with automated data analysis.


2020 ◽  
Vol 86 (12) ◽  
Author(s):  
Christian Keggi ◽  
Joy Doran-Peterson

ABSTRACT Paenibacillus amylolyticus 27C64, a Gram-positive bacterium with diverse plant cell wall polysaccharide deconstruction capabilities, was isolated previously from an insect hindgut. Previous work suggested that this organism’s pectin deconstruction system differs from known systems in that its sole pectin methylesterase is cytoplasmic, not extracellular. In this work, we have characterized the specific roles of key extracellular pectinases involved in homogalacturonan deconstruction, including four pectate lyases and one pectin lyase. We show that one newly characterized pectate lyase, PelC, has a novel substrate specificity, with a lower Km for highly methylated pectins than for polygalacturonic acid. PelC works synergistically with PelB, a high-turnover exo-pectate lyase that releases Δ4,5-unsaturated trigalacturonate as its major product. It is likely that PelC frees internal stretches of demethylated homogalacturonan which PelB can degrade. We also show that the sole pectin lyase has a high kcat value and rapidly depolymerizes methylated substrates. Three cytoplasmic GH105 hydrolases were screened for the ability to remove terminal unsaturated galacturonic acid residues from oligogalacturonide products produced by the action of extracellular lyases, and we found that two are active on demethylated oligogalacturonides. This work confirms that efficient homogalacturonan deconstruction in P. amylolyticus 27C65 does not require extracellular pectin methylesterase activity. Three of the extracellular lyases studied in this work are also thermostable, function well over a broad pH range, and have significant industrial potential. IMPORTANCE Pectin is an important structural polysaccharide found in most plant cell walls. In the environment, pectin degradation is part of the decomposition process that turns over dead plant material and is important to organisms that feed on plants. Industrially, pectinases are used to improve the quality of fruit juices and can also be used to process coffee cherries or tea leaves. These enzymes may also prove useful in reducing the environmental impact of paper and cotton manufacturing. This work is significant because it focuses on a Gram-positive bacterium that is evolutionarily distinct from other well-studied pectin-degrading organisms and differs from known systems in key ways. Most importantly, a simplified extracellular deconstruction process in this organism is able to break down pectins without first removing the methyl groups that inhibit other systems. Moreover, some of the enzymes described here have the potential to improve industrial processes that rely on pectin deconstruction.


2018 ◽  
Vol 200 (18) ◽  
Author(s):  
James A. Budnick ◽  
Lauren M. Sheehan ◽  
Lin Kang ◽  
Pawel Michalak ◽  
Clayton C. Caswell

ABSTRACTElucidating the function of proteins <50 amino acids in length is no small task. Nevertheless, small proteins can play vital roles in the lifestyle of bacteria and influence the virulence of pathogens; thus, the investigation of the small proteome is warranted. Recently, our group identified theBrucella abortusprotein VtlR as a transcriptional activator of four genes, one of which is the well-studied small regulatory RNA AbcR2, while the other three genes encode hypothetical small proteins, two of which are highly conserved among the orderRhizobiales. This study provides evidence that all three genes encode authentic small proteins and that all three are highly expressed under oxidative stress, low-pH, and stationary-phase growth conditions. Fractionation of the cells revealed that the proteins are localized to the membranes ofB. abortus. We demonstrate that the small proteins under the transcriptional control of VtlR are not accountable for attenuation observed with theB. abortusvtlRdeletion strain. However, there is an association between VtlR-regulated genes and growth inhibition in the presence of the sugarl-fucose. Subsequent transcriptomic analyses revealed thatB. abortusinitiates the transcription of a locus encoding a putative sugar transport and utilization system when the bacteria are cultured in the presence ofl-fucose. Altogether, our observations characterize the role of the VtlR-controlled small proteins BAB1_0914, BAB2_0512, and BAB2_0574 in the biology ofB. abortus, particularly in the capacity of the bacteria to utilizel-fucose.IMPORTANCEDespite being one of the most common zoonoses worldwide, there is currently no human vaccine to combat brucellosis. Therefore, a better understanding of the pathogenesis and biology ofBrucellaspp., the causative agent of brucellosis, is essential for the discovery of novel therapeutics against these highly infectious bacteria. In this study, we further characterize the virulence-associated transcriptional regulator VtlR inBrucella abortus. Our findings not only shed light on our current understanding of a virulence related genetic system inBrucellaspp. but also increase our knowledge of small proteins in the field of bacteriology.


2014 ◽  
Vol 70 (a1) ◽  
pp. C749-C749
Author(s):  
Kunio Yubuta ◽  
Yusuke Mizuno ◽  
Nobuyuki Zettsu ◽  
Shigeki Komine ◽  
Kenichiro Kami ◽  
...  

Present spinel-type lithium manganese oxides have attracted much attention as positive-electrode active materials for lithium-ion rechargeable batteries, which are the most sought-after power source for various electric applications, because of their low cost, non-toxicity, and high abundance of source materials compared to the conventionally used LiCoO2 crystals. Spinel-type LiMn2O4 crystals were grown at low-temperature by using a LiCl-KCl flux. The chemical compositions, sizes, and shapes of the LiMn2O4 crystals could be tuned by simply changing the growth conditions. Among the various products, the crystals grown at a low temperature of 873 K showed a small average size of 200 nm. Electron diffraction patterns and TEM images reveal the truncated octahedral shape of the crystals. The flux growth driven by rapid cooling resulted in truncated octahedral LiMn2O4 crystals surrounded by both dominating {111} and minor {100} faces with {311} and {220} edges. Lattice images indicate that crystals grown at a lower temperature have the excellent crystallinity. The small LiMn2O4 crystals grown at 873 K showed better rate properties than the large crystals grown at 1173 K, when used as a positive active material in lithium-ion rechargeable batteries.


Sign in / Sign up

Export Citation Format

Share Document