{Reference Type}: Journal Article
{Author}: Sygmund, C.; Gutmann, A.; Krondorfer, I.; Kujawa, M.; Glieder, A.; Pscheidt, B.; Haltrich, D.; Peterbauer, C.; Kittl, R.
{Year}: 2012
{Title}: Simple and efficient expression of Agaricus meleagris pyranose dehydrogenase in Pichia pastoris
{URL}: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=22080342&query_hl=1
{Tag}: 0
{Star}: 0
{Journal}: Appl Microbiol Biotechnol
{Volume}: 94
{Issue}: 3
{Pages}: 695-704
{DOI}: 10.1007/s00253-011-3667-7
{Date Displayed}: 2012 May
{Date}: 2012-05-01
{Type of Work}: Journal Article; Research Support, Non-U.S. Gov't
{Accession Number}: 22080342
{Keywords}: Agaricus/*enzymology/genetics; *Carbohydrate Metabolism; *Gene Expression; Oxidoreductases/*biosynthesis/genetics; Pichia/*genetics; Recombinant Proteins/biosynthesis/genetics
{Abstract}: Pyranose dehydrogenase (PDH) is a fungal flavin-dependent sugar oxidoreductase that is highly interesting for applications in organic synthesis or electrochemistry. The low expression levels of the filamentous fungus Agaricus meleagris as well as the demand for engineered PDH make heterologous expression necessary. Recently, Aspergillus species were described to efficiently secrete recombinant PDH. Here, we evaluate recombinant protein production with expression hosts more suitable for genetic engineering. Expression in Escherichia coli resulted in no soluble or active PDH. Heterologous expression in the methylotrophic yeast Pichia pastoris was investigated using two different signal   sequences as well as a codon-optimized sequence. A 96-well plate activity screening for transformants of all constructs was established and the best expressing clone was used for large-scale production in 50-L scale, which gave a   volumetric yield of 223 mg L(-1) PDH or 1,330 U L(-1) d(-1) in space-time yield.   Purification yielded 13.4 g of pure enzyme representing 95.8% of the initial activity. The hyperglycosylated recombinant enzyme had a 20% lower specific activity than the native enzyme; however, the kinetic properties were essentially identical. This study demonstrates the successful expression of PDH in the eukaryotic host organism P. pastoris paving the way for protein engineering. Additionally, the feasibility of large-scale production of the enzyme with this expression system together with a simplified purification scheme for easy high-yield purification is shown.
{Author Address}: Department of Food Sciences and Technology, University of Natural Resources and Applied Life Sciences (BOKU), Muthgasse 11, 1190 Vienna, Austria.
{Language}: eng

{Reference Type}: Journal Article
{Author}: Zhang, L.; Pei, Y.; Xing, Z.; Ding, S.; Buswell, J. A.
{Year}: 2011
{Title}: Comparison of endoglucanase-1 (EG1) induction in the edible straw mushroom Volvariella volvacea by lactose and/or cellobiose with or without added sorbose
{URL}: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=21076917&query_hl=1
{Tag}: 0
{Star}: 0
{Journal}: Appl Microbiol Biotechnol
{Volume}: 89
{Issue}: 6
{Pages}: 1939-46
{DOI}: 10.1007/s00253-010-2995-3
{Date Displayed}: 2011 Mar
{Date}: 2011-03-01
{Type of Work}: Journal Article; Research Support, Non-U.S. Gov't
{Accession Number}: 21076917
{Keywords}: Cellobiose/*metabolism; Cellulase/*biosynthesis; Culture Media/chemistry; Gene Expression Profiling; *Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Fungal; Lactose/*metabolism; Reverse Transcriptase Polymerase Chain Reaction; Sorbose/*metabolism; Time Factors; Volvariella/*enzymology
{Abstract}: We have compared the induction of an endoglucanase (EG1) by alpha-lactose and/or   cellobiose, with or without added L-sorbose, in submerged cultures of Volvariella volvacea, to better understand the mechanism whereby cellulase formation is triggered by these soluble disaccharides. EG1 levels induced by alpha-lactose and cellobiose were 28.6% and 6.7%, respectively of the highest levels recorded with   crystalline cellulose. Sorbose did not induce EG1 and strongly repressed enzyme levels when added to alpha-lactose but not cellobiose-containing cultures. EG1 levels in cultures containing all three saccharides were similar to those recorded with sorbose and cellobiose although enzyme induction was delayed by 12   h. When V. volvacea was pre-grown for 24 h in medium containing sorbose as the sole carbon source, followed by addition of alpha-lactose or cellobiose or a mixture of the two, EG1 levels recorded in the alpha-lactose-supplemented cultures were again markedly lower compared with cultures containing only alpha-lactose. Maximal enzyme levels in cultures with added cellobiose or cellobiose and alpha-lactose were not affected although appearance of EG1 in culture supernatants was again delayed by 12 h. Semi-quantitative RT-PCR revealed that higher, more prolonged, levels of eg1 transcription occurred in V. volvacea   cultures induced with alpha-lactose compared with cellobiose- or alpha-lactose +   cellobiose-induced cultures. However, eg1 transcription levels in cultures induced with cellobiose or with cellobiose + lactose, and the corresponding cultures with added sorbose, were not markedly different.
{Author Address}: Department of Biological Engineering, Nanjing Forestry University, Nanjing, Jiangsu, People's Republic of China.
{Language}: eng

{Reference Type}: Journal Article
{Author}: Largeteau, M. L.; Llarena-Hernandez, R. C.; Regnault-Roger, C.; Savoie, J. M.
{Year}: 2011
{Title}: The medicinal Agaricus mushroom cultivated in Brazil: biology, cultivation and non-medicinal valorisation
{URL}: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=22005742&query_hl=1
{Tag}: 0
{Star}: 0
{Journal}: Appl Microbiol Biotechnol
{Volume}: 92
{Issue}: 5
{Pages}: 897-907
{DOI}: 10.1007/s00253-011-3630-7
{Date Displayed}: 2011 Dec
{Date}: 2011-12-01
{Type of Work}: Journal Article; Research Support, Non-U.S. Gov't; Review
{Accession Number}: 22005742
{Keywords}: Agaricus/*chemistry/enzymology/genetics/*growth & development; Brazil; Culture Techniques/*methods; Functional Food/*analysis; Fungal Proteins/genetics/metabolism; Nutritive Value; Plant Diseases/*prevention & control
{Abstract}: Sun mushroom is a cultivated mushroom extensively studied for its medicinal properties for several years and literature abounds on the topic. Besides, agronomical aspects were investigated in Brazil, the country the mushroom comes from, and some studies focus on the biology of the fungus. This review aimed to present an overview of the non-medicinal knowledge on the mushroom. Areas of commercial production and marketing trends are presented. Its specific fragrance, taste, nutritional value and potential use of extracts as food additives are compared to those of the most cultivated fungi and laboratory models. The interest of the mushroom for lignocellulosic enzyme production and source of biomolecules for the control of plant pathogens are shown. Investigation of genetic variability among cultivars is reported. Growing and storage of mycelium, as well as cultivation conditions (substrate and casing generally based on local   products; indoor and outdoor cultivation; diseases and disorders) are described and compared to knowledge on Agaricus bisporus.
{Author Address}: INRA, UR1264, MycSA, 33883, Villenave d'Ornon, France. largeteau@bordeaux.inra.fr
{Language}: eng

{Reference Type}: Journal Article
{Author}: Singh, R. K.; Zhang, Y. W.; Nguyen, N. P.; Jeya, M.; Lee, J. K.
{Year}: 2011
{Title}: Covalent immobilization of beta-1,4-glucosidase from Agaricus arvensis onto functionalized silicon oxide nanoparticles
{URL}: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=20811797&query_hl=1
{Tag}: 0
{Star}: 0
{Journal}: Appl Microbiol Biotechnol
{Volume}: 89
{Issue}: 2
{Pages}: 337-44
{DOI}: 10.1007/s00253-010-2768-z
{Date Displayed}: 2011 Jan
{Date}: 2011-01-01
{Type of Work}: Journal Article; Research Support, Non-U.S. Gov't
{Accession Number}: 20811797
{Keywords}: Agaricus/chemistry/*enzymology/genetics/isolation & purification; Enzyme Stability; Enzymes, Immobilized/*chemistry/isolation & purification; Fungal Proteins/*chemistry/isolation & purification; Kinetics; Molecular Weight; Nanoparticles/*chemistry; Silicon Dioxide/chemistry; Soil Microbiology; beta-Glucosidase/*chemistry/isolation & purification
{Abstract}: An efficient beta-1,4-glucosidase (BGL) secreting strain, Agaricus arvensis, was   isolated and identified. The relative molecular weight of the purified A. arvensis BGL was 98 kDa, as determined by sodium dodecylsulfate polyacrylamide gel electrophoresis, or 780 kDa by size exclusion chromatography, indicating that the enzyme is an octamer. Using a crude enzyme preparation, A. arvensis BGL was covalently immobilized onto functionalized silicon oxide nanoparticles with an immobilization efficiency of 158%. The apparent V (max) (k (cat)) values of free   and immobilized BGL under standard assay conditions were 3,028 U mg protein(-1) (4,945 s(-1)) and 3,347 U mg protein(-1) (5,466 s(-1)), respectively. The immobilized BGL showed a higher optimum temperature and improved thermostability   as compared to the free enzyme. The half-life at 65 degrees C showed a 288-fold improvement over the free BGL. After 25 cycles, the immobilized enzyme still retained 95% of the original activity, thus demonstrating its prospects for commercial applications. High specific activity, high immobilization efficiency,   improved stability, and reusability of A. arvensis BGL make this enzyme of potential interest in a number of industrial applications.
{Author Address}: Department of Chemical Engineering, Konkuk University, 1 Hwayang-Dong, Gwangjin-Gu, Seoul, South Korea, 143-701.
{Language}: eng