Stability of hydrolytic enzymes in water-organic solvent systems

Simon, Lajos; László, K.; Vértesi, Adél; Bagi, K.; Szajáni, B.

doi:10.1016/s1381-1177(97)00019-2

Cited by 60 publications

(32 citation statements)

References 14 publications

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“…The lipase produced by A. mediterranei DSM 43304 was remarkably stable in both hydrophilic and hydrophobic organic solvents (Table 6). Hydrophilic solvents are generally incompatible with enzyme activity because they remove the crucial bound water from the enzyme surface, thereby destabilizing the protein and causing high denaturation rates [47,60]. Interestingly, the lipase from A. mediterranei DSM 43304 presented an opposite behavior and showed activation with solvents of log P values less than -0.2.…”

Section: Discussionmentioning

confidence: 99%

Influence of cultivation conditions on the production of a thermostable extracellular lipase from Amycolatopsis mediterranei DSM 43304

Dheeman¹,

Frias²,

Henehan³

2009

J Ind Microbiol Biotechnol

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23Among several lipase producing actinomycete strains screened, Amycolatopsis 24 mediterranei DSM 43304 was found to produce a thermostable, extracellular lipase. 25Culture condition and nutrient source modification studies involving carbon sources, 26 nitrogen sources, incubation temperature and medium pH were carried out. Lipase activity 27 of 1.37 ± 0.103 IU/ml of culture medium was obtained in 96 h at 28 °C and pH 7.5 using 28 linseed oil and fructose as carbon sources and a combination of phytone peptone and yeast 29 extract (5:1) as nitrogen sources. In optimal culture conditions the lipase activity was 30 enhanced 12-fold with a 2-fold increase in lipase specific activity. The lipase showed 31 maximum activity at 60 °C and pH 8.

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Section: Discussionmentioning

confidence: 99%

Influence of cultivation conditions on the production of a thermostable extracellular lipase from Amycolatopsis mediterranei DSM 43304

Dheeman¹,

Frias²,

Henehan³

2009

J Ind Microbiol Biotechnol

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“…Consequently, many efforts have been devoted to overcoming the low solubility of sterols and improving product yield. These include the application of various water-miscible organic solvents (Simon et al, 1998), water-immiscible organic solvents (Cruz et al, 2001), silicone oil (Kutney et al, 2000;Stefanov et al, 2006), polypropylene glycol (Kutney et al, 2000;Stefanov et al, 2006), cloud point system (Wang et al, 2004;, and microemulsions (Stefan et al, 2002). However, organic solvents present many drawbacks such as toxicity to microbial cells and environmental hazards (Schimid et al, 1998;Kim et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

Optimization of biotransformation from phytosterol to androstenedione by a mutant Mycobacterium neoaurum ZJUVN-08

Zhang

Peng

et al. 2013

J. Zhejiang Univ. Sci. B

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Biotransformation of phytosterol (PS) by a newly isolated mutant Mycobacterium neoaurum ZJUVN-08 to produce androstenedione has been investigated in this paper. The parameters of the biotransformation process were optimized using fractional factorial design and response surface methodology. Androstenedione was the sole product in the fermentation broth catalyzed by the mutant M. neoaurum ZJUVN-08 strain. Results showed that molar ratio of hydroxypropyl-β-cyclodextrin (HP-β-CD) to PS and substrate concentrations were the two most significant factors affecting androstenedione production. By analyzing the statistical model of three-dimensional surface plot, the optimal process conditions were observed at 0.1 g/L inducer, pH 7.0, molar ratio of HP-β-CD to PS 1.92:1, 8.98 g/L PS, and at 120 h of incubation time. Under these conditions, the maximum androstenedione yield was 5.96 g/L and nearly the same with the non-optimized (5.99 g/L), while the maximum PS conversion rate was 94.69% which increased by 10.66% compared with the non-optimized (84.03%). The predicted optimum conditions from the mathematical model were in agreement with the verification experimental results. It is considered that response surface methodology was a powerful and efficient method to optimize the parameters of PS biotransformation process.

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“…As shown in Figure 2b, pre-incubation of CT in ACN for 10 min (CT blank 10 min) prior to the 5 min activity assay (CT blank) lowers the activity from 100 to 50%. This can be attributed to the detrimental influence of 50% ACN on CT activity, as has previously been reported for other enzyme systems (Griebenow and Klibanov, 1996;Simon et al, 1998;Olofsson et al, 2005). In contrast to the studies performed in buffer, neither the presence of lysates 1-4 nor the reference proteins during the 10 min pre-incubation affected CT activity significantly.…”

Section: Resultsmentioning

confidence: 49%

Phage display screening in low dielectric media

Olofsson¹,

Söderberg²,

Ankarloo³

et al. 2008

J of Molecular Recognition

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Here we report the first application of phage display screening in low dielectric media. Two series of phage clones with affinity for alpha-chymotrypsin (CT) were selected from a Ph.D.(TM)-C7C library, using either a buffer or acetonitrile in buffer (50%, v/v). The affinity of lysates, individual clones or selected cyclic peptides for the enzyme was studied by examining their influence on CT activity. Peptides displayed on phage selected in buffer provided significant protection from enzyme autolysis resulting in marked increase in CT activity (>100%). Phage selected in ACN provided some, albeit weak, protection from the detrimental influence on CT from ACN. In conclusion, the results demonstrate the potential for the application of phage display screening protocols to targets in media of low dielectricity.

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Stability of hydrolytic enzymes in water-organic solvent systems

Cited by 60 publications

References 14 publications

Influence of cultivation conditions on the production of a thermostable extracellular lipase from Amycolatopsis mediterranei DSM 43304

Influence of cultivation conditions on the production of a thermostable extracellular lipase from Amycolatopsis mediterranei DSM 43304

Optimization of biotransformation from phytosterol to androstenedione by a mutant Mycobacterium neoaurum ZJUVN-08

Phage display screening in low dielectric media

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