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https://www.readbyqxmd.com/read/28811204/heterologous-expression-and-biochemical-characterization-of-a-novel-thermostable-sclerotinia-sclerotiorum-gh45-endoglucanase-in-pichia-pastoris
#1
Haifa Chahed, Mohamed Boumaiza, Aymen Ezzine, M Nejib Marzouki
Enzymatic saccharification of lignocellulosic biomass has been widely studied. Mainly endoglucanases were found to be a prerequisite for the quick initial biomass liquefaction. In the present study, Pichia pastoris was used as a host for the heterologous expression of a Sclerotinia sclerotiorum GH45 endoglucanase, Endo2. The recombinant plasmid pPICZαA was used to transform Pichia pastoris. Pichia culture supernatants expressing the recombinant Endo2 (rEndo2) were used for the purification and biochemical characterization of this enzyme...
August 12, 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/28807840/the-heterologous-expression-strategies-of-antimicrobial-peptides-in-microbial-systems
#2
REVIEW
Ting Deng, Haoran Ge, Huahua He, Yao Liu, Chao Zhai, Liang Feng, Li Yi
Antimicrobial peptides (AMPs) consist of molecules acting on the defense systems of numerous organisms toward tumor and multiple pathogens, such as bacteria, fungi, viruses, and parasites. Compared to traditional antibiotics, AMPs are more stable and have lower propensity for developing resistance through functioning in the innate immune system, thus having important applications in the fields of medicine, food and so on. However, despite of their high economic values, the low yield and the cumbersome extraction process in AMPs production are problems that limit their industrial application and scientific research...
August 12, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28807829/the-magnaporthe-oryzae-alt-a-1-like-protein-mohrip1-binds-to-the-plant-plasma-membrane
#3
Yi Zhang, Yingbo Liang, Yijie Dong, Yuhan Gao, Xiufen Yang, Jingjing Yuan, Dewen Qiu
MoHrip1, a protein isolated from Magnaporthe oryzae, belongs to the Alt A 1 (AA1) family. mohrip1 mRNA levels showed inducible expression throughout the infection process in rice. To determine the location of MoHrip1 in M. oryzae, a mohrip1-gfp mutant was generated. Fluorescence microscopy observations and western blotting analysis showed that MoHrip1 was both present in the secretome and abundant in the fungal cell wall. To obtain MoHrip1 protein, we carried out high-yield expression of MoHrip1 in Pichia pastoris...
August 12, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28807724/the-versatile-functions-of-lrr-only-proteins-in-mollusk-chlamys-farreri
#4
Mengqiang Wang, Lingling Wang, Zhihao Jia, Xiudan Wang, Qilin Yi, Zhao Lv, Linsheng Song
Leucine-rich repeat (LRR)-only proteins are involved in the innate immune responses as they mediate protein-ligand interactions. In the present study, three novel LRR-only proteins, CfLRRop-4, CfLRRop-5 and CfLRRop-6, were identified and characterized from Zhikong scallop Chlamys farreri. They all contained LRR motifs with consensus signature sequences of LxxLxLxxNxL or LxxLxLxxCxxL. All the mRNA transcripts of three CfLRRops were high abundant in hepatopancreas, gills and gonads, and their mRNA transcripts in hemocytes could respond to the stimulations of different microbes, including Vibrio anguillarum, Micrococcus luteus and Pichia pastoris...
August 11, 2017: Developmental and Comparative Immunology
https://www.readbyqxmd.com/read/28807601/ubx-domain-containing-proteins-are-involved-in-lipid-homeostasis-and-stress-responses-in-pichia-pastoris
#5
Meng Zhang, Qilin Yu, Zhe Liu, Chen Liang, Biao Zhang, Mingchun Li
Ubiquitin regulatory X (UBX) domain-containing proteins constitute a family of proteins and are substrate adaptors of AAA ATPase Cdc48. UBX proteins can bind to the N-terminal region of Cdc48 to perform endoplasmic reticulum associated protein degradation (ERAD). In this study, we identified two UBX domain-containing proteins, Ubx1 and Ubx2, in Pichia pastoris and found that the two proteins could recover the growth defect of Saccharomyces cerevisiae in ubx2Δ. Our results revealed that Ubx1 and Ubx2 play critical roles in synthesis of unsaturated fatty acids by affecting Spt23...
August 12, 2017: International Journal of Biochemistry & Cell Biology
https://www.readbyqxmd.com/read/28800907/biochemical-characterization-of-aspergillus-oryzae-recombinant-%C3%AE-l-rhamnosidase-expressed-in-pichia-pastoris
#6
Mai Ishikawa, Yoshihito Shiono, Takuya Koseki
An α-l-rhamnosidase-encoding gene from Aspergillus oryzae, which belongs to the glycoside hydrolase family 78, was cloned and expressed in Pichia pastoris. SDS-PAGE of the purified recombinant α-l-rhamnosidase protein revealed smeared bands with apparent molecular mass of 90-130 kDa. After N-deglycosylation, the recombinant enzyme showed a molecular mass of 70 kDa. The enzyme exhibited optimal activity at a pH of 5.0 and a temperature of 70 °C. Specific activity of the enzyme was higher toward hesperidin than toward naringin, which consist of α-1,6 and α-1,2 linkages, respectively...
August 9, 2017: Journal of Bioscience and Bioengineering
https://www.readbyqxmd.com/read/28797224/the-adaptive-landscape-of-wildtype-and-glycosylation-deficient-populations-of-the-industrial-yeast-pichia-pastoris
#7
Josef W Moser, Iain B H Wilson, Martin Dragosits
BACKGROUND: The effects of long-term environmental adaptation and the implications of major cellular malfunctions are still poorly understood for non-model but biotechnologically relevant species. In this study we performed a large-scale laboratory evolution experiment with 48 populations of the yeast Pichia pastoris in order to establish a general adaptive landscape upon long-term selection in several glucose-based growth environments. As a model for a cellular malfunction the implications of OCH1 mannosyltransferase knockout-mediated glycosylation-deficiency were analyzed...
August 10, 2017: BMC Genomics
https://www.readbyqxmd.com/read/28779880/investigation-of-a-nosocomial-outbreak-of%C3%A2-fungemia-caused-by-candida-pelliculosa-pichia-anomala-in-a-korean-tertiary-care-center
#8
Jiwon Jung, Young Sook Moon, Jung A Yoo, Ji-Hun Lim, Joseph Jeong, Jae-Bum Jun
BACKGROUND: Candida pelliculosa is a rare pathogen of fungemia. There have been a few nosocomial outbreaks of C. pelliculosa fungemia in nurseries and pediatric intensive care units (ICU), hematologic units, and surgical ICU. We describe an epidemiologic outbreak investigation, including case findings of C. pelliculosa fungemia in South Korea. METHODS: This outbreak investigation conducted in a 940-bed, tertiary referral center, Ulsan, South Korea and included active microbial surveillance and a case-control study...
June 28, 2017: Journal of Microbiology, Immunology, and Infection, Wei Mian Yu Gan Ran za Zhi
https://www.readbyqxmd.com/read/28776697/an-automated-data-driven-dsp-development-approach-for-glycoproteins-from-yeast
#9
Vignesh Rajamanickam, Maximillian Krippl, Christoph Herwig, Oliver Spadiut
Downstream process development for recombinant glycoproteins from yeast is cumbersome due to hyperglycosylation of target proteins. In a previous study, we purified three recombinant glycoproteins from Pichia pastoris using a simple two-step flowthrough mode approach using monolithic columns. In this study, we investigated a novel automated data science approach for identifying purification conditions for such glycoproteins using monolithic columns. We performed three sets of design of experiments (DoEs) in analytical scale to determine the separation efficiency of monolithic columns for three different recombinant horseradish peroxidase (HRP) isoenzymes...
August 4, 2017: Electrophoresis
https://www.readbyqxmd.com/read/28771577/a-survey-of-the-mycobiota-associated-with-larvae-of-the-black-soldier-fly-hermetia-illucens-reared-for-feed-production
#10
Ilaria Varotto Boccazzi, Matteo Ottoboni, Elena Martin, Francesco Comandatore, Lisa Vallone, Thomas Spranghers, Mia Eeckhout, Valeria Mereghetti, Luciano Pinotti, Sara Epis
Feed security, feed quality and issues surrounding the safety of raw materials are always of interest to all livestock farmers, feed manufacturers and competent authorities. These concerns are even more important when alternative feed ingredients, new product developments and innovative feeding trends, like insect-meals, are considered. The black soldier fly (Hermetia illucens) is considered a good candidate to be used as feed ingredient for aquaculture and other farm animals, mainly as an alternative protein source...
2017: PloS One
https://www.readbyqxmd.com/read/28771492/production-of-an-anti-a%C3%AE-antibody-fragment-in-pichia-pastoris-and-in-vitro-and-in-vivo-validation-of-its-therapeutic-effect
#11
Laia Montoliu-Gaya, Gisela Esquerda-Canals, Silvia Bronsoms, Sandra Villegas
ScFv-h3D6 has been shown as an efficient therapy in the 3xTg-AD mouse model of Alzheimer's Disease. Because one of the major bottlenecks for the therapeutic uses of proteins produced in Escherichia coli is their potential contamination with endotoxins, LPS were extensively removed by a rather low-efficient, expensive, and time-consuming purification step. In addition, disulfide scrambling is favored in the reducing bacterial cytoplasm albeit the use of reductase deficient strains. To overcome these hurdles, as well as to improve the yield, the yeast Pichia pastoris, an endotoxin-free host system for recombinant protein production, has been used to produce scFv-h3D6, both in flask and in a fed-batch bioreactor...
2017: PloS One
https://www.readbyqxmd.com/read/28769888/unraveling-core-functional-microbiota-in-traditional-solid-state-fermentation-by-high-throughput-amplicons-and-metatranscriptomics-sequencing
#12
Zhewei Song, Hai Du, Yan Zhang, Yan Xu
Fermentation microbiota is specific microorganisms that generate different types of metabolites in many productions. In traditional solid-state fermentation, the structural composition and functional capacity of the core microbiota determine the quality and quantity of products. As a typical example of food fermentation, Chinese Maotai-flavor liquor production involves a complex of various microorganisms and a wide variety of metabolites. However, the microbial succession and functional shift of the core microbiota in this traditional food fermentation remain unclear...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28764098/two-acidic-thermophilic-gh28-polygalacturonases-from-talaromyces-leycettanus-jcm-12802-with-application-potentials-for-grape-juice-clarification
#13
Yeqing Li, Yuan Wang, Tao Tu, Duoduo Zhang, Rui Ma, Shuai You, Xiaoyu Wang, Bin Yao, Huiying Luo, Bo Xu
Efficient hydrolysis of pectic materials to sugars requires the synergistic action of endo- and exo-polygalacturonases. Two novel polygalacturonases (exo-TePG28a and endo-TePG28b) were identified in Talaromyces leycettanus JCM12802, overexpressed in Pichia pastoris, and characterized in this report. The specific activities of TePG28a and TePG28b towards polygalacturonic acid were 280±9 and 25,900±502U/mg, respectively. Both enzymes exhibited optimal activities at pH 3.5 and retained highly stable over a broad pH range of 2...
December 15, 2017: Food Chemistry
https://www.readbyqxmd.com/read/28764073/an-improved-process-for-the-production-of-highly-purified-recombinant-thaumatin-tagged-variants
#14
Robert D Healey, Helene Lebhar, Simon Hornung, Pall Thordarson, Christopher P Marquis
The sweetest tasting molecule known is the protein thaumatin, first isolated from the katemfe fruit, Thaumatococcus daniellii. Thaumatin is used in the food and beverage industry as a low-calorie sugar substitute. Thaumatin interacts with taste receptors in the oral cavity eliciting a persistent sweet taste and a bitter, liquorice flavor. Recombinant thaumatin was expressed in Pichia pastoris and through a co-expression strategy with a molecular chaperone, yields of one engineered thaumatin variant increased by greater than two-fold...
December 15, 2017: Food Chemistry
https://www.readbyqxmd.com/read/28764054/seed-incorporation-during-vinification-and-its-impact-on-chemical-and-organoleptic-properties-in-syzygium-cumini-wine
#15
K S VenuGopal, K A Anu-Appaiah
Syzgium cumini (Jamun) is an evergreen tropical tree, its various parts are known for many therapeutic properties. The present work represents the production of wines from jamun fruits using two different native isolates (Saccharomyces cerevisiae - KF551990 and Pichia gummiguttae - MCC 1273) and influence of jamun seeds on the physico-chemical parameters, chromatic properties, phenolic components and sensory attributes of wine. Wine produced was bottle aged for one year. On aging there was a reduction in bitterness and astringency...
December 15, 2017: Food Chemistry
https://www.readbyqxmd.com/read/28763110/co-expression-of-lipase-isozymes-for-enhanced-expression-in-pichia-pastoris
#16
H Cai, T Zhang, M Zhao, J Mao, C Cai, F Feng
To enhance the expression level of overall lipase isozymes which catalyze the same reaction in P. pastoris through co-expression of isozymes from different sources, several types of co-expression ways were constructed to determine the co-expression efficiencies of lipase isozymes in P. pastoris. The results showed that the Kex2 mediated co-expression of lipase isozymes could express Rhizomucor miehei lipase (RML) and Thermomyces lanuginosus lipase (TLL) simultaneously, and GS-RMk-kTL displayed an average lipase activity of 306...
August 1, 2017: Letters in Applied Microbiology
https://www.readbyqxmd.com/read/28762562/co-expression-of-alpha-1-antitrypsin-with-cytoplasmic-domain-of-v-snare-in-pichia-pastoris-preserving-biological-activity-of-alpha-1-antitrypsin
#17
Maryam Khatami, Seyed Nezamedin Hosseini, Sadegh Hasannia
Alpha-1-antitrypsin (A1AT) is a major serum protein in human with protease inhibitory activity. Because of its extensive application in medicine, recombinant DNA technology has been considered for its production. The current study is going to examine co-expression of A1AT, and soluble domain of v-SNARE in Pichia pastoris, which can prevent the secretion of A1AT after passing the secretory pathway thoroughly. This was done mainly to preserve the biological activity of A1AT which in the secretory mode might be impaired in the fermentation and early clarification conditions...
August 1, 2017: Biotechnology and Applied Biochemistry
https://www.readbyqxmd.com/read/28761999/expression-of-chondroitin-4-o-sulfotransferase-in-escherichia-coli-and-pichia-pastoris
#18
Wenqin He, Yuanyuan Zhu, Abhijeet Shirke, Xiaojun Sun, Jian Liu, Richard A Gross, Mattheos A G Koffas, Robert J Linhardt, Ming Li
Chondroitin sulfates are linear sulfated polysaccharides called glycosaminoglycans. They are important nutraceutical and pharmaceutical products that are biosynthesized through the action of chondroitin sulfotransferases on either an unsulfated chondroitin or a dermatan polysaccharide precursor. While the enzymes involved in the biosynthesis of chondroitin sulfates are well known, the cloning end expression of these membrane-bound Golgi enzymes continue to pose challenges. The major chondroitin-4-sulfotransferase, Homo sapiens C4ST-1, had been previously cloned and expressed from mammalian CHO, COS-7, and HEK 293 cells, and its activity was shown to require glycosylation...
July 31, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28760369/towards-systems-metabolic-engineering-in-pichia-pastoris
#19
REVIEW
Jan-Philipp Schwarzhans, Tobias Luttermann, Martina Geier, Jörn Kalinowski, Karl Friehs
The methylotrophic yeast Pichia pastoris is firmly established as a host for the production of recombinant proteins, frequently outperforming other heterologous hosts. Already, a sizeable amount of systems biology knowledge has been acquired for this non-conventional yeast. By applying various omics-technologies, productivity features have been thoroughly analyzed and optimized via genetic engineering. However, challenging clonal variability, limited vector repertoire and insufficient genome annotation have hampered further developments...
July 29, 2017: Biotechnology Advances
https://www.readbyqxmd.com/read/28759578/heterologous-expression-of-the-antimyotoxic-protein-dm64-in-pichia-pastoris
#20
Saulo Martins Vieira, Surza Lucia Gonçalves da Rocha, Ana Gisele da Costa Neves-Ferreira, Rodrigo Volcan Almeida, Jonas Perales
Snakebite envenomation is a neglected condition that constitutes a public health problem in tropical and subtropical countries, including Brazil. Interestingly, some animals are resistant to snake envenomation due to the presence of inhibitory glycoproteins in their serum that target toxic venom components. DM64 is an acidic glycoprotein isolated from Didelphis aurita (opossum) serum that has been characterized as an inhibitor of the myotoxicity induced by bothropic toxins bearing phospholipase A2 (PLA2) structures...
July 2017: PLoS Neglected Tropical Diseases
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