Purification and characterization of α-glucosidase from Aspergillus carbonarious

Summary An -glucosidase was purified from Aspergillus carbonarious CCRC 30414 over 20 fold with 37 % recovery. Its molecular mass was estimated to be 328 kDa by gel filtration with an optimum pH from 4.2 to 5.0, and pI=5.0. The optimum temperature is at 60°C over 40 min. The enzyme was partially inhibited by 5 mM Ag⁺, Hg²⁺, Ba²⁺, Pb²⁺, and Aso₄ ⁺.     

Immobilization of <Emphasis Type="Bold">β</Emphasis>-fructofuranosidase from <Emphasis Type="Italic">Aspergillus japonicus</Emphasis> on chitosan using tris(hydroxymethyl)phosphine or glutaraldehyde as a coupling agent

A partially purified -fructofuranosidase from Aspergillus japonicus was covalently immobilized on to chitosan beads using either glutaraldehyde or tris(hydroxymethyl)phosphine (THP) as a coupling agent. Compared with the glutaraldehyde-immobilized and the free enzyme, the THP-immobilized enzyme had the highest thermal stability with 78% activity retained after 12 days at 37 ° C. The THP-immobilized enzyme also had higher reusability than that immobilized by glutaraldehyde, 75% activity was retained after 11 batches (or 11 days) at 37° C for the THP immobilized enzyme system. Less yield (48%) of fructooligosaccharides (FOS) were produced by the THP-immobilized enzyme compared with the free enzyme system (58%) from 50 (w/v) sucrose at 50 ° C.     

Cyclodextrin Glucanotransferase from Alkalophilic Bacteria of Taiwan

Chloromonilinic acids A and B were newly isolated from a culture filtrate of M. fructicola accumutating chloromonilicin in the mycelium, and their structures were elucidated on the basis of spectro scopic evidence. The unusual catabolic route of chloromonilicin is discussed.     

A peroxiredoxin cDNA from Taiwanofungus camphorata: role of Cys31 in dimerization

Peroxiredoxins (Prxs) play important roles in antioxidant defense and redox signaling pathways. A Prx isozyme cDNA (TcPrx2, 745 bp, EF552425) was cloned from Taiwanofungus camphorata and its recombinant protein was overexpressed. The purified protein was shown to exist predominantly as a dimer by sodium dodecyl sulfate-polyacrylamide gel electrolysis in the absence of a reducing agent. The protein in its dimeric form showed no detectable Prx activity. However, the protein showed increased Prx activity with increasing dithiothreitol concentration which correlates with dissociation of the dimer into monomer. The TcPrx2 contains two Cys residues. The Cys⁶⁰ located in the conserved active site is the putative active peroxidatic Cys. The role of Cys³¹ was investigated by site-directed mutagenesis. The C31S mutant (C³¹ → S³¹) exists predominantly as a monomer with noticeable Prx activity. The Prx activity of the mutant was higher than that of the corresponding wild-type protein by nearly twofold at 12 μg/mL. The substrate preference of the mutant was H₂O₂ > cumene peroxide > t-butyl peroxide. The Michaelis constant (K _M) value for H₂O₂ of the mutant was 0.11 mM. The mutant enzyme was active under a broad pH range from 6 to 10. The results suggest a role of Cys³¹ in dimerization of the TcPrx2, a role which, at least in part, may be involved in determining the activity of Prx. The C³¹ residue does not function as a resolving Cys and therefore the TcPrx2 must follow the reaction mechanism of 1-Cys Prx. This TcPrx2 represents a new isoform of Prx family.     

Production of High-content Galacto-oligosaccharide by Enzyme Catalysis and Fermentation with Kluyveromyces marxianus

Of three β-galactosidases from Aspergillus oryzae, Kluyveromyces lactis and Bacillus sp., used for the production of low-content galacto- oligosaccharides (GOS) from lactose, the latter produced the highest yield of trisaccharides and tetrasaccharides. GOS production was enhanced by mixing β-galactosidase glucose oxidase. The low-content GOS syrups, produced either by β-galactosidase alone or by the mixed enzyme system, were subjected to the fermentation by Kluyveromyces marxianus, whereby glucose, galactose, lactose and other disaccharides were depleted, resulting in up to 97% and 98% on a dry weight basis of high-content GOS with the yields of 31% and 32%, respectively. An erratum to this article can be found at     

Continuous production of high-content fructooligosaccharides by a complex cell system

A complex biocatalyst system with a bioreactor equipped with a microfiltration (MF) module was employed to produce high-content fructooligosaccharides (FOS) in a continuous process initiated by a batch process. The system used mycelia of Aspergillus japonicus CCRC 93007 or Aureobasidium pullulans ATCC 9348 with beta-fructofuranosidase activity and Gluconobacter oxydans ATCC 23771 with glucose dehydrogenase activity. Calcium carbonate slurry was used to control pH to 5.5, and gluconic acid in the reaction mixture was precipitated as calcium gluconate. Sucrose solution with an optimum concentration of 30% (w/v) was employed as feed for the complex cell system, and high-content FOS was discharged continuously from a MF module. The complex cell system was run at 30 degrees C with an aeration rate of 5 vvm and produced more than 80% FOS with the remainder being 5-7% glucose and 8-10% sucrose on a dry weight basis, plus a small amount of calcium gluconate. The system worked for a 7-day continuous production process with a dilution rate of 0.04 h(-1), and the volumetric productivity for total FOS was more than 160 g L(-1) h(-1).     

Production of xylitol from Candida tropicalis by using an oxidation-reduction potential-stat controlled fermentation

An on-line device, ORP (oxidation-reduction potential)-stat, was used to control glucose-feeding for enhancing xylitol conversion from D-xylose during an oxygen-limited fermentation by Candida tropicalis. The fermentation was carried out in a 5 l jar fermenter. After glucose in the medium was depleted, a switching to a limited aeration and feeding glucose controlled by ORP-stat was performed. The maximum xylitol yield was obtained under a condition at an ORP of -180 mV and at an aeration rate of 0.2 l min(-1).     

Production of xylitol from Candida tropicalis by using an oxidation-reduction potential-stat controlled fermentation

An on-line device, ORP (oxidation-reduction potential)-stat, was used to control glucose-feeding for enhancing xylitol conversion from D-xylose during an oxygen-limited fermentation by Candida tropicalis. The fermentation was carried out in a 5 l jar fermenter. After glucose in the medium was depleted, a switching to a limited aeration and feeding glucose controlled by ORP-stat was performed. The maximum xylitol yield was obtained under a condition at an ORP of - 180 mV and at an aeration rate of 0.2 l min(-1).