Extranuclear Expression of the Bacterial Xylose Isomerase ( xyl A) and the UDP-Glucose Dehydrogenase ( has B) Genes in Yeast with Kluyveromyces lactis Linear Killer Plasmids as Vectors

1996 ◽  
Vol 33 (5) ◽  
pp. 323-330 ◽  
Author(s):  
Jürgen Schründer ◽  
Norio Gunge ◽  
Friedhelm Meinhardt
Keyword(s):  
Kluyveromyces Lactis ◽  
Glucose Dehydrogenase ◽  
Xylose Isomerase

scholarly journals Heterologous Hyaluronic Acid Production in Kluyveromyces lactis

2019 ◽  
Vol 7 (9) ◽  
pp. 294
Author(s):  
Antonio M. V. Gomes ◽  
João H. C. M. Netto ◽  
Lucas S. Carvalho ◽  
Nádia S. Parachin
Keyword(s):  
Hyaluronic Acid ◽  
Pathogenic Bacteria ◽  
Pasteurella Multocida ◽  
Glucuronic Acid ◽  
Kluyveromyces Lactis ◽  
Glucose Dehydrogenase ◽  
Primary Sources ◽  
Batch Mode ◽  
Dehydrogenase Gene ◽  
Ha Synthase

Hyaluronic Acid (HA) is a biopolymer composed by the monomers Glucuronic Acid (GlcUA) and N-Acetyl Glucosamine (GlcNAc). It has a broad range of applications in the field of medicine, being marketed between USD 1000–5000/kg. Its primary sources include extraction of animal tissue and fermentation using pathogenic bacteria. However, in both cases, extensive purification protocols are required to prevent toxin contamination. In this study, aiming at creating a safe HA producing microorganism, the generally regarded as safe (GRAS) yeast Kluyveroymyces lactis is utilized. Initially, the hasB (UDP-Glucose dehydrogenase) gene from Xenopus laevis (xlhasB) is inserted. After that, four strains are constructed harboring different hasA (HA Synthase) genes, three of humans (hshasA1, hshasA2, and hshasA3) and one with the bacteria Pasteurella multocida (pmhasA). Transcript values analysis confirms the presence of hasA genes only in three strains. HA production is verified by scanning electron microscopy in the strain containing the pmHAS isoform. The pmHAS strain is grown in a 1.3 l bioreactor operating in a batch mode, the maximum HA levels are 1.89 g/L with a molecular weight of 2.097 MDa. This is the first study that reports HA production in K. lactis and it has the highest HA titers reported among yeast.

scholarly journals Engineering Pseudomonas putida S12 for Efficient Utilization of d-Xylose and l-Arabinose

2008 ◽  
Vol 74 (16) ◽  
pp. 5031-5037 ◽  
Author(s):  
Jean-Paul Meijnen ◽  
Johannes H. de Winde ◽  
Harald J. Ruijssenaars
Keyword(s):  
Growth Rate ◽  
Pseudomonas Putida ◽  
Glucose Dehydrogenase ◽  
Xylose Isomerase ◽  
Dry Weight ◽  
High Yield ◽  
Efficient Production ◽  
Primary Metabolism ◽  
Dead End ◽  
Solvent Tolerant

ABSTRACT The solvent-tolerant bacterium Pseudomonas putida S12 was engineered to utilize xylose as a substrate by expressing xylose isomerase (XylA) and xylulokinase (XylB) from Escherichia coli. The initial yield on xylose was low (9% [g CDW g substrate−1], where CDW is cell dry weight), and the growth rate was poor (0.01 h−1). The main cause of the low yield was the oxidation of xylose into the dead-end product xylonate by endogenous glucose dehydrogenase (Gcd). Subjecting the XylAB-expressing P. putida S12 to laboratory evolution yielded a strain that efficiently utilized xylose (yield, 52% [g CDW g xylose−1]) at a considerably improved growth rate (0.35 h−1). The high yield could be attributed in part to Gcd inactivity, whereas the improved growth rate may be connected to alterations in the primary metabolism. Surprisingly, without any further engineering, the evolved d-xylose-utilizing strain metabolized l-arabinose as efficiently as d-xylose. Furthermore, despite the loss of Gcd activity, the ability to utilize glucose was not affected. Thus, a P. putida S12-derived strain was obtained that efficiently utilizes the three main sugars present in lignocellulosic hydrolysate: glucose, xylose, and arabinose. This strain will form the basis for a platform host for the efficient production of biochemicals from renewable feedstock.

scholarly journals Refactoring the upper sugar metabolism ofPseudomonas putidafor co-utilization of disaccharides, pentoses, and hexoses

2018 ◽  
Author(s):  
Pavel Dvořák ◽  
Víctor de Lorenzo
Keyword(s):  
Glucose Dehydrogenase ◽  
Sugar Metabolism ◽  
Xylose Isomerase ◽  
Pentose Phosphate ◽  
Soil Bacterium ◽  
Biochemical Network ◽  
Xylose Metabolism ◽  
Native Proteins ◽  
Dead End ◽  
A Genome

AbstractGiven its capacity to tolerate stress, NAD(P)H/ NAD(P) balance, and increased ATP levels, the platform strainPseudomonas putidaEM42, a genome-edited derivative of the soil bacteriumP. putidaKT2440, can efficiently host a suite of harsh reactions of biotechnological interest. Because of the lifestyle of the original isolate, however, the nutritional repertoire ofP. putidaEM42 is centered largely on organic acids, aromatic compounds and some hexoses (glucose and fructose). To enlarge the biochemical network ofP. putidaEM42 to include disaccharides and pentoses, we implanted heterologous genetic modules for D-cellobiose and D-xylose metabolism into the enzymatic complement of this strain. Cellobiose was actively transported into the cells through the ABC complex formed by native proteins PP1015-PP1018. The knocked-in β-glucosidase BglC fromThermobifida fuscacatalyzed intracellular cleavage of the disaccharide to D-glucose, which was then channelled to the default central metabolism. Xylose oxidation to the dead end product D-xylonate was prevented by by deleting thegcdgene that encodes the broad substrate range quinone-dependent glucose dehydrogenase. Intracellular intake was then engineered by expressing theEscherichia coliproton-coupled symporter XylE. The sugar was further metabolized by the products ofE. coli xylA(xylose isomerase) andxylB(xylulokinase) towards the pentose phosphate pathway. The resultingP. putidastrain co-utilized xylose with glucose or cellobiose to complete depletion of the sugars. These results not only show the broadening of the metabolic capacity of a soil bacterium towards new substrates, but also promoteP. putidaEM42 as a platform for plug-in of new biochemical pathways for utilization and valorization of carbohydrate mixtures from lignocellulose processing.

scholarly journals The Mitochondrial Genome Integrity Gene, MGI1, of Kluyveromyces lactis Encodes the β-Subunit of F1-ATPase

Genetics ◽  
1996 ◽  
Vol 144 (4) ◽  
pp. 1445-1454 ◽  
Author(s):  
Xin Jie Chen ◽  
G Desmond Clark-Walker
Keyword(s):  
Mitochondrial Genome ◽  
Kluyveromyces Lactis ◽  
Three Dimensional ◽  
Genome Integrity ◽  
Dimensional Structure ◽  
Deletion Mutants ◽  
Β Subunit ◽  
Gain Of Function ◽  
F1 Atpase ◽  
Γ Subunit

In a previous report, we found that mutations at the mitochondrial genome integrity locus, MGI1, can convert Kluyveromyces lactis into a petite-positive yeast. In this report, we describe the isolation of the MGI1 gene and show that it encodes the β-subunit of the mitochondrial F1-ATPase. The site of mutation in four independently isolated mgi1 alleles is at Arg435, which has changed to Gly in three cases and Ile in the fourth isolate. Disruption of MGI1 does not lead to the production of mitochondrial genome deletion mutants, indicating that an assembled F1 complex is needed for the “gain-of-function” phenotype found in mgi1 point mutants. The location of Arg435 in the β-subunit, as deduced from the three-dimensional structure of the bovine F1-ATPase, together with mutational sites in the previously identified mgi2 and mgi5 alleles, suggests that interaction of the β- and α- (MGI2) subunits with the γ-subunit (MGI5) is likely to be affected by the mutations.

scholarly journals Towards a Novel Computer-Aided Optimization of Microreactors: Techno-Economic Evaluation of an Immobilized Enzyme System

Symmetry ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 524
Author(s):  
Philip Pietrek ◽  
Manfred Kraut ◽  
Roland Dittmeyer
Keyword(s):  
Magnetic Beads ◽  
Glucose Dehydrogenase ◽  
Packed Bed ◽  
Detailed Knowledge ◽  
Reaction Conditions ◽  
Flow Reactors ◽  
Continuous Flow Reactors ◽  
Enzyme Cascades ◽  
And Control ◽  
Physical And Chemical

Immobilized multi-enzyme cascades are increasingly used in microfluidic devices. In particular, their application in continuous flow reactors shows great potential, utilizing the benefits of reusability and control of the reaction conditions. However, capitalizing on this potential is challenging and requires detailed knowledge of the investigated system. Here, we show the application of computational methods for optimization with multi-level reactor design (MLRD) methodology based on the underlying physical and chemical processes. We optimize a stereoselective reduction of a diketone catalyzed by ketoreductase (Gre2) and Nicotinamidadenindinukleotidphosphat (NADPH) cofactor regeneration with glucose dehydrogenase (GDH). Both enzymes are separately immobilized on magnetic beads forming a packed bed within the microreactor. We derive optimal reactor feed concentrations and enzyme ratios for enhanced performance and a basic economic model in order to maximize the techno-economic performance (TEP) for the first reduction of 5-nitrononane-2,8-dione.

Preparation, characterization, and in vivo evaluation of anti-inflammatory activities of selenium nanoparticles synthesized by Kluyveromyces lactis GG799

2021 ◽  
Author(s):  
Xiao fan Song ◽  
Lei Qiao ◽  
Shuqi Yan ◽  
Yue Chen ◽  
Xina Dou ◽  
...  
Keyword(s):  
Inflammatory Bowel Disease ◽  
Bowel Disease ◽  
Kluyveromyces Lactis ◽  
Selenium Nanoparticles ◽  
Human Diseases ◽  
In Vivo Evaluation ◽  
Inflammatory Bowel ◽  
Anti Inflammatory

Selenium (Se) as an essential micronutrient that has implications in human diseases, including inflammatory bowel disease (IBD), especially with respect to Se deficiencies. Recently, selenium nanoparticles (SeNPs) have attracted significant...

Sign in / Sign up

Export Citation Format

Share Document