Process development for high-level secretory production of carboxypeptidase Y by Saccharomyces cerevisiae

1998 ◽  
Vol 50 (1) ◽  
pp. 34-41 ◽  
Author(s):  
Y. Shiba ◽  
F. Fukui ◽  
K. Ichikawa ◽  
N. Serizawa ◽  
H. Yoshikawa
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Process Development ◽  
Carboxypeptidase Y ◽  
Secretory Production ◽  
High Level

scholarly journals High-Level Secretory Production of Phospholipase A1by Saccharomyces cerevisiae and Aspergillus oryzae

2001 ◽  
Vol 65 (1) ◽  
pp. 94-101 ◽  
Author(s):  
Yoichiro SHIBA ◽  
Chiho ONO ◽  
Fumio FUKUI ◽  
Ichiro WATANABE ◽  
Nobufusa SERIZAWA ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Aspergillus Oryzae ◽  
Secretory Production ◽  
High Level

High-level secretory production of recombinant human lipocortin-I by Saccharomyces cerevisiae

1999 ◽  
Vol 35 (1-2) ◽  
pp. 97-101 ◽  
Author(s):  
Bong Hyun Chung ◽  
Dong Jin Seo ◽  
Soo Wan Nam
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Secretory Production ◽  
High Level

scholarly journals Tempo and Mode of Ty Element Evolution in Saccharomyces cerevisiae

Genetics ◽  
1999 ◽  
Vol 151 (4) ◽  
pp. 1341-1351 ◽  
Author(s):  
I King Jordan ◽  
John F McDonald
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Negative Selection ◽  
Sequence Variation ◽  
Size Variation ◽  
Sequence Comparisons ◽  
Ty Elements ◽  
Ty Element ◽  
Amino Acid Sequence Variation ◽  
Element Evolution ◽  
High Level

Abstract The Saccharomyces cerevisiae genome contains five families of long terminal repeat (LTR) retrotransposons, Ty1–Ty5. The sequencing of the S. cerevisiae genome provides an unprecedented opportunity to examine the patterns of molecular variation existing among the entire genomic complement of Ty retrotransposons. We report the results of an analysis of the nucleotide and amino acid sequence variation within and between the five Ty element families of the S. cerevisiae genome. Our results indicate that individual Ty element families tend to be highly homogenous in both sequence and size variation. Comparisons of within-element 5′ and 3′ LTR sequences indicate that the vast majority of Ty elements have recently transposed. Furthermore, intrafamily Ty sequence comparisons reveal the action of negative selection on Ty element coding sequences. These results taken together suggest that there is a high level of genomic turnover of S. cerevisiae Ty elements, which is presumably in response to selective pressure to escape host-mediated repression and elimination mechanisms.

Functional domains of SIR4, a gene required for position effect regulation in Saccharomyces cerevisiae

1987 ◽  
Vol 7 (12) ◽  
pp. 4441-4452
Author(s):  
M Marshall ◽  
D Mahoney ◽  
A Rose ◽  
J B Hicks ◽  
J R Broach
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Mating Type ◽  
Position Effect ◽  
Protein Activity ◽  
Multicomponent Complex ◽  
Mating Type Genes ◽  
Chromosome Iii ◽  
Covalently Linked ◽  
High Level ◽  
Mat Locus

The product of the Saccharomyces cerevisiae SIR4 gene, in conjunction with at least three other gene products, prevents expression of mating-type genes resident at loci at either end of chromosome III, but not of the same genes resident at the MAT locus in the middle of the chromosome. To address the mechanism of this novel position effect regulation, we have conducted a structural and genetic analysis of the SIR4 gene. We have determined the nucleotide sequence of the gene and found that it encodes a lysine-rich, serine-rich protein of 152 kilodaltons. Expression of the carboxy half of the protein complements a chromosomal nonsense mutation of sir4 but not a complete deletion of the gene. These results suggest that SIR4 protein activity resides in two portions of the molecule, but that these domains need not be covalently linked to execute their biological function. We also found that high-level expression of the carboxy domain of the protein yields dominant derepression of the silent loci. This anti-Sir activity can be reversed by increased expression of the SIR3 gene, whose product is normally also required for maintaining repression of the silent loci. These results are consistent with the hypothesis that SIR3 and SIR4 proteins physically associate to form a multicomponent complex required for repression of the silent mating-type loci.

Establishment of a Competitive Additive Manufacturing Workshop

2016 ◽  
Author(s):  
Paul Ryan ◽  
Jan Schwerdtfeger ◽  
Markus Rodermann
Keyword(s):  
Additive Manufacturing ◽  
Gas Turbines ◽  
High Performance ◽  
Best Practice ◽  
Process Development ◽  
Process Efficiency ◽  
Development Effort ◽  
Industrial Gas Turbines ◽  
High Level ◽  
Design And Manufacture

Compared to conventional manufacturing processes, additive manufacturing offers a degree of freedom that has the potential to revolutionize the turbine components supply chain. Additive manufacturing facilitates the design and manufacture of highly complex components in high performance materials with features that cannot currently be realized with other processes. In addition, shorter development and manufacturing lead-times are possible due to the flexibility of 3D based processing and the absence of expensive, complicated molds or dies. Having been confined for many years to rapid prototyping or R&D applications, additive manufacturing is now making the move to the factory floor. However, a dearth of manufacturing experience makes the development effort and risk of costly mistakes a deterrent for many organizations that would otherwise be interested in exploring the benefits of additive manufacturing. A former manufacturer of industrial gas turbines recently established an additive manufacturing workshop designed to deliver highly complex engine-ready components that can contribute to increased performance of the gas turbine. A strong emphasis on process validation and implementation of the organization’s best practice Lean and Quality methodologies has laid solid foundations for a highly capable manufacturing environment. This paper describes the approach taken to ensure that the workshop achieves a high level of operational excellence. Process development topics explored in the paper include the following: • Planning of process flow and cell layout to permit the maximum lean performance • Strategy used to determine machine specification and selection method. • Assessment of process capability • Influence of design for manufacture on process efficiency and product quality • Experience gained during actual production of first commercial components

scholarly journals Isolation and characterization of PEP3, a gene required for vacuolar biogenesis in Saccharomyces cerevisiae

1991 ◽  
Vol 11 (12) ◽  
pp. 5801-5812
Author(s):  
R A Preston ◽  
M F Manolson ◽  
K Becherer ◽  
E Weidenhammer ◽  
D Kirkpatrick ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Gene Product ◽  
Zinc Finger ◽  
Genomic Library ◽  
Sequence Similarity ◽  
Carboxypeptidase Y ◽  
Lag Phase ◽  
Significant Similarity ◽  
Reading Frame ◽  
Isolation And Characterization

The Saccharomyces cerevisiae PEP3 gene was cloned from a wild-type genomic library by complementation of the carboxypeptidase Y deficiency in a pep3-12 strain. Subclone complementation results localized the PEP3 gene to a 3.8-kb DNA fragment. The DNA sequence of the fragment was determined; a 2,754-bp open reading frame predicts that the PEP3 gene product is a hydrophilic, 107-kDa protein that has no significant similarity to any known protein. The PEP3 predicted protein has a zinc finger (CX2CX13CX2C) near its C terminus that has spacing and slight sequence similarity to the adenovirus E1a zinc finger. A radiolabeled PEP3 DNA probe hybridized to an RNA transcript of 3.1 kb in extracts of log-phase and diauxic lag-phase cells. Cells bearing pep3 deletion/disruption alleles were viable, had decreased levels of protease A, protease B, and carboxypeptidase Y antigens, had decreased repressible alkaline phosphatase activity, and contained very few normal vacuolelike organelles by fluorescence microscopy and electron microscopy but had an abundance of extremely small vesicles that stained with carboxyfluorescein diacetate, were severely inhibited for growth at 37 degrees C, and were incapable of sporulating (as homozygotes). Fractionation of cells expressing a bifunctional PEP3::SUC2 fusion protein indicated that the PEP3 gene product is present at low abundance in both log-phase and stationary cells and is a vacuolar peripheral membrane protein. Sequence identity established that PEP3 and VPS18 (J. S. Robinson, T. R. Graham, and S. D. Emr, Mol. Cell. Biol. 11:5813-5824, 1991) are the same gene.

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