46 research outputs found
Thermal inactivation and conformational lock studies on glucose oxidase
In this study, the dissociative thermal inactivation
and conformational lock theories are applied for the
homodimeric enzyme glucose oxidase (GOD) in order to
analyze its structure. For this purpose, the rate of activity
reduction of glucose oxidase is studied at various temperatures
using b-D-glucose as the substrate by incubation of
enzyme at various temperatures in the wide range between
40 and 70 �C using UV–Vis spectrophotometry. It was
observed that in the two ranges of temperatures, the
enzyme has two different forms. In relatively low temperatures,
the enzyme is in its dimeric state and has normal
activity. In high temperatures, the activity almost disappears
and it aggregates. The above achievements are confirmed
by dynamic light scattering. The experimental
parameter ‘‘n’’ as the obvious number of conformational
locks at the dimer interface of glucose oxidase is obtained
by kinetic data, and the value is near to two. To confirm the
above results, the X-ray crystallography structure of the
enzyme, GOD (pdb, 1gal), was also studied. The secondary
and tertiary structures of the enzyme to track the thermal
inactivation were studied by circular dichroism and
fluorescence spectroscopy, respectively. We proposed a
mechanism model for thermal inactivation of GOD based
on the absence of the monomeric form of the enzyme by
circular dichroism and fluorescence spectroscopy
A Novel Strategy to Construct Yeast Saccharomyces cerevisiae Strains for Very High Gravity Fermentation
Very high gravity (VHG) fermentation is aimed to considerably increase both the fermentation rate and the ethanol concentration, thereby reducing capital costs and the risk of bacterial contamination. This process results in critical issues, such as adverse stress factors (ie., osmotic pressure and ethanol inhibition) and high concentrations of metabolic byproducts which are difficult to overcome by a single breeding method. In the present paper, a novel strategy that combines metabolic engineering and genome shuffling to circumvent these limitations and improve the bioethanol production performance of Saccharomyces cerevisiae strains under VHG conditions was developed. First, in strain Z5, which performed better than other widely used industrial strains, the gene GPD2 encoding glycerol 3-phosphate dehydrogenase was deleted, resulting in a mutant (Z5ΔGPD2) with a lower glycerol yield and poor ethanol productivity. Second, strain Z5ΔGPD2 was subjected to three rounds of genome shuffling to improve its VHG fermentation performance, and the best performing strain SZ3-1 was obtained. Results showed that strain SZ3-1 not only produced less glycerol, but also increased the ethanol yield by up to 8% compared with the parent strain Z5. Further analysis suggested that the improved ethanol yield in strain SZ3-1 was mainly contributed by the enhanced ethanol tolerance of the strain. The differences in ethanol tolerance between strains Z5 and SZ3-1 were closely associated with the cell membrane fatty acid compositions and intracellular trehalose concentrations. Finally, genome rearrangements in the optimized strain were confirmed by karyotype analysis. Hence, a combination of genome shuffling and metabolic engineering is an efficient approach for the rapid improvement of yeast strains for desirable industrial phenotypes
PRODUCTION OF LIGNOLYTIC AND FEED-BACK TYPE ENZYMES BY PHLEBIA RADIATA ON DIFFERENT MEDIA
Low molecular-weight compounds, structurally related to lignin, increase the production of laccase, lignin peroxidase, manganese dependent peroxidase, and feed-back type enzymes such as glucose oxi- dase, cellobioso-quinone oxidoreductase, and glyoxal oxidase in the culture of the white rot fungus Phlebia radiata growing on different carbon sources
Low-temperature biocatalysis as an example of microbial biotransformation of alpha-pinene
Katalizatory warunkują przebieg oraz odpowiednią wydajność wielu reakcji chemicznych. Przemiany biochemiczne w większości przypadków katalizowane są z udziałem enzymów. Szczególne cechy biokatalizatorów są powszechnie wykorzystywane w przemyśle, a w ostatnich latach drobnoustroje ekstremofilne, adaptowane do skrajnych warunków abiotycznych, wzbudzają duże zainteresowanie licznej grupy badaczy. Wyjątkowe predyspozycje biokatalizatora psychrotroficznego z dobrymi efektami zostały wykorzystane w procesie biotransformacji a-pinenu do werbenolu i werbenonu, będących cennymi związkami smakowo-zapachowymi stosowanymi w branży spożywczej i kosmetycznej. Uzyskane wyniki (sumaryczne stężenie produktów przekraczające 0,5 g/dm3) stanowią obiecującą perspektywę wykorzystania tego typu procesów w większej skali (ponadlaboratoryjnej).Many chemical reactions and their appropriate performance depend on catalysis. Most biochemical reactions are catalysed by enzymes. Special features of these biocatalysts are commonly employed in industry and current researches are focused on psychrotrophic microorganisms. These unique organisms can be a rich source of useful biocatalysts for biotransformation of a-pinene to verbenol and verbenone - valuable compounds used as flavours and fragrances. Results obtained in this study are promising for further application in the large scale
Izolacja i wstepna charakterystyka niskoproteazowych mutantow Trichoderma reesei
Po mutagenizacji promieniowaniem UV i NTG szczepu Trichoderma reesei M-7 i selekcji na podłożu z dodatkiem żelatyny wyizolowano około 960 niskoproteazowych mutantów. Dziesięć z nich zostało przetestowanych podczas hodowli okresowych w zakresie produkcji celulaz, proteaz i beta-galaktozydazy w obecności serwatki i laktozy jako źródła węgla. Aktywności proteolityczne oznaczone po hodowlach 9 mutantów w obecności laktozy były niższe od szczepu wyjściowego. Zaobserwowano większy indukcyjny wpływ serwatki niż laktozy, na sekrecję proteaz przez wszystkie badane szczepy. Czysta laktoza okazała się bardziej efektywnym induktorem produkcji beta-galaktozydazy niż zawarta w serwatce. Plony biomasy większości mutantów po hodowlach na serwatce były tylko nieznacznie wyższe od oznaczonych po hodowlach na czystej laktozie. Było to prawdopodobnie związane z litycznym mechanizmem kompensującym proteaz, przejawiających wyższe aktywności w filtratach pochodzących z hodowli w obecności serwatki. Praca jest wstępem do dalszych badań nad zależnościami pomiędzy proteazami i innymi enzymami w filtratach pohodowlanych Trichoderma reesei.It has been isolated 960 low-protease mutants after mutagenisation using UV radiation and NTG and selection on medium with addition of gellatine. Ten of them have been tested during batch cultivation for the production of cellulases, proteases and beta-galactosidase in the presence of whey and lactose as a source of carbon. Proteolytic activities estimated after cultivation of 9 mutants in the presence of lactose were lower then parental strain. It has been observed better inductive effect of whey then lactose on the secretion of proteases by all mutants tested. Pure lactose appeared to be more effective inducer of the production of beta-galactosidase then contains in whey. Biomass concentration of most mutants after cultivation on whey was only slightly higher then estimated after cultivation on pure lactose. It was probably connected with lytic compensatory mechanism of proteases exhibited higher activities in culture filtrates from cultivation in the presence of whey. This work is an introduction for future studies on correlation between proteases and other enzymes in culture filtrates of Trichoderma reesei
Mimetic host shifts in an endangered social parasite of ants
An emerging problem in conservation is whether listed morpho-species with broad distributions, yet specialized lifestyles, consist of more than one cryptic species or functionally distinct forms that have different ecological requirements. We describe extreme regional divergence within an iconic endangered butterfly, whose socially parasitic young stages use non-visual, non-tactile cues to infiltrate
and supplant the brood in ant societies. Although indistinguishable morphologically or when using current mitochondrial and nuclear sequence-, or microsatellite data, Maculinea rebeli from Spain and southeast Poland exploit
different Myrmica ant species and experience 100 per cent mortality with each other’s hosts. This reflects major differences in the hydrocarbons synthesized from each region by the larvae, which so closely mimic the recognition profiles of their respective hosts that nurse ants afford each parasite a social status above that of their own kin larvae. The two host ants occupy separate niches within grassland; thus, conservation management must differ in each region. Similar cryptic differentiation may be common, yet equally hard to detect, among the approximately 10 000 unstudied morpho-species of social parasite that are estimated to exist, many of which are Red Data Book listed