Stabilization of d-amino acid oxidase from Rhodosporidium toruloides by immobilization onto magnetic nanoparticles

Hsieh, Hao‐Chieh; Kuan, I-Ching; Lee, Shiow-Ling; Tien, Gee-Yeng; Wang, Yi-Jen; Yu, Chih-Wen

doi:10.1007/s10529-008-9894-z

Cited by 25 publications

(11 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This loss is likely to be a result of the dissolution of the Ca‐alginate matrix, which will lead to a loss of the entrapped enzyme and consequently, a decrease in the catalytic activity. The fact that entrapped F54Y can be used for 20 cycles of reaction is a substantial improvement when compared with a previous attempt in which covalently immobilized DAAO lost more than half of its activity after only five reaction cycles 12. Although comparable results have been reported in another study of immobilized Tv DAAO 14, a comparison of the protocols reveals that our immobilization method was fast, simple to operate with higher yield, and required no hazardous reagent (glutaraldehyde).…”

Section: Resultssupporting

confidence: 63%

“…Covalent binding is a common method of immobilization. Although improved enzyme performance and operational stability were observed in previous studies on the covalent immobilization of Tv DAAO, the procedures were tedious and involved the use of toxic cross‐linking reagents 11–14. In addition, covalent methods may also cause alterations in protein conformation and yield inactive immobilizates 15.…”

Section: Introductionmentioning

confidence: 99%

“…In addition, covalent methods may also cause alterations in protein conformation and yield inactive immobilizates 15. Thus, the covalently immobilized Tv DAAO exhibits reduced cofactor affinity 11 and low operational stability, as reflected by a significant loss of catalytic activity (>50%) after five reaction cycles 12. On the other hand, enzyme entrapment and encapsulation are considered as mild immobilization methods with minimal disturbance in protein structure and catalytic activity 15, 16.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Entrapment of a Trigonopsis variabilis D‐amino acid oxidase variant F54Y for oxidative deamination of cephalosporin C

Wong¹,

Fong²,

Tsang³

2011

Engineering in Life Sciences

View full text Add to dashboard Cite

Trigonopsis variabilis D‐amino acid oxidase (TvDAAO) is an enzyme used in the industrial bioconversion of cephalosporin C (CPC) into 7‐aminocephalosporanic acid, a crucial biosynthetic nucleus for a wide spectrum of semi‐synthetic cephem antibiotics. Using homology modeling and site‐directed mutagenesis, we have previously shown that the TvDAAO variant F54Y possesses improved catalytic activity and thermostability. To further explore its industrial application, the conditions for immobilization of the enzyme were examined in the present investigation. The results showed that entrapment in a calcium alginate (Ca‐alginate) matrix using 2% alginate, 500 mM CaCl2, and 15 min stabilization appeared to be optimal for the immobilization of F54Y. The entrapped enzyme allowed complete CPC conversion. The entrapped enzyme also showed good operational stability and retained at least 90% of its original activity after 20 reaction cycles. To conclude, the entrapment of F54Y in Ca‐alginate appeared to be a simple and efficient biocatalysis system with potential application in the antibiotics industry.

show abstract

Section: Resultssupporting

confidence: 63%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Entrapment of a Trigonopsis variabilis D‐amino acid oxidase variant F54Y for oxidative deamination of cephalosporin C

Wong¹,

Fong²,

Tsang³

2011

Engineering in Life Sciences

View full text Add to dashboard Cite

show abstract

“…In the presence of 50 mM hydrogen peroxide, encapsulated RtDAO had approximately 2-fold increase in resistance to hydrogen peroxide. Glutyaraldehyde immobilization on magnetic nanoparticles of the same enzyme also coupled thermal stabilization (Tm was increased from 45 °C to 55 °C) with stabilization in the presence of 20 mM H 2 O 2 , (the immobilized form retained 93% of its activity after 5 h while the free form was completely inactivated after 3.5 h) [254]. Multipoint covalently immobilized D-amino acid oxidase (DAAO) from Rhodotorula gracilis on glyoxyl-agarose is 11-fold more stable than the native enzyme against the deleterious effect of hydrogen peroxide, with an improved thermostabillity as an added benefit [251,255].…”

Section: Improved Rigidity Of the Enzymementioning

confidence: 99%

Hydrogen Peroxide in Biocatalysis. A Dangerous Liaison

Hernández

Berenguer-Murcia²,

Rodrigues³

et al. 2012

COC

140

107

View full text Add to dashboard Cite

Hydrogen peroxide is a substrate or side-product in many enzyme-catalyzed reactions. For example, it is a side-product of oxidases, resulting from the re-oxidation of FAD with molecular oxygen, and it is a substrate for peroxidases and other enzymes. However, hydrogen peroxide is able to chemically modify the peptide core of the enzymes it interacts with, and also to produce the oxidation of some cofactors and prostetic groups (e.g., the hemo group). Thus, the development of strategies that may permit to increase the stability of enzymes in the presence of this deleterious reagent is an interesting target. This enhancement in enzyme stability has been attempted following almost all available strategies: site-directed mutagenesis (eliminating the most reactive moieties), medium engineering (using stabilizers), immobilization and chemical modification (trying to generate hydrophobic environments surrounding the enzyme, to confer higher rigidity to the protein or to generate oxidation-resistant groups), or the use of systems capable of decomposing hydrogen peroxide under very mild conditions. If hydrogen peroxide is just a side-product, its immediate removal has been reported to be the best solution. In some cases, when hydrogen peroxide is the substrate and its decomposition is not a sensible solution, researchers coupled one enzyme generating hydrogen peroxide "in situ" to the target enzyme resulting in a continuous supply of this reagent at low concentrations thus preventing enzyme inactivation.This review will focus on the general role of hydrogen peroxide in biocatalysis, the main mechanisms of enzyme inactivation produced by this reactive and the different strategies used to prevent enzyme inactivation caused by this "dangerous liaison". Outlook

show abstract

“…The oleaginous yeast Rhodosporidium toruloides is a versatile yeast that can produce biocatalysts, amino acids and lipid (Hsieh et al, 2009;Jia et al, 2008). R. toruloides Y4 was an excellent storage lipid producer because high cellular lipid content (Li et al, 2006) and high cell density (Li et al, 2007) could be easily achieved.…”

Section: Introductionmentioning

confidence: 99%

Microbial lipid production by Rhodosporidium toruloides under sulfate-limited conditions

Zhao

Shen

et al. 2011

Bioresource Technology

193

102

View full text Add to dashboard Cite

Stabilization of d-amino acid oxidase from Rhodosporidium toruloides by immobilization onto magnetic nanoparticles

Cited by 25 publications

References 22 publications

Entrapment of a Trigonopsis variabilis D‐amino acid oxidase variant F54Y for oxidative deamination of cephalosporin C

Entrapment of a Trigonopsis variabilis D‐amino acid oxidase variant F54Y for oxidative deamination of cephalosporin C

Hydrogen Peroxide in Biocatalysis. A Dangerous Liaison

Microbial lipid production by Rhodosporidium toruloides under sulfate-limited conditions

Contact Info

Product

Resources

About