Controlled water content for evaluation of denaturation temperature of freeze-dried enzymes

Videa, Marcelo; Cruz-Angeles, Jorge; Rodríguez, Sara Luisa

doi:10.1016/j.tca.2016.06.013

Cited by 7 publications

(3 citation statements)

References 33 publications

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“…Peak melting from thermograms vs hydration ratio h for a series of samples at different scan rates. Continuous and dashed lines are from global fitting of literature data, average and confidence bands, respectively. Data obtained at a scan rate of 10 K/min are from this work (blue solid triangles), ref (black open squares), ref (red solid stars), and ref (black solid squares).…”

Section: Resultsmentioning

confidence: 99%

Specific Interactions and Environment Flexibility Tune Protein Stability under Extreme Crowding

et al. 2021

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Macromolecular crowding influences protein mobility and stability in vivo. A precise description of the crowding effect on protein thermal stability requires the estimate of the combined effects of excluded volume, specific protein−environment interactions, as well as the thermal response of the crowders. Here, we explore an ideal model system, the lysozyme protein in powder state, to dissect the factors controlling the melting of the protein under extreme crowding. By deploying state-of-the art molecular simulations, supported by calorimetric experiments, we assess the role of the environment flexibility and of intermolecular electrostatic interactions. In particular, we show that the temperature-dependent flexibility of the macromolecular crowders, along with specific interactions, significantly alleviates the stabilizing contributions of the static volume effect.

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Section: Resultsmentioning

confidence: 99%

Specific Interactions and Environment Flexibility Tune Protein Stability under Extreme Crowding

et al. 2021

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“…Previous studies suggested that the presence of excess water facilitated irreversible denaturation of immobilized lipases by promoting conformational mobility of protein molecules and hydrolysis of peptide bonds . A reduced water content around protein molecules modifies the hydrogen bond interactions between water and protein, thereby increasing the denaturation temperature . In addition, the application of vacuum provided a relatively inert atmosphere which may enhance the thermostability of enzymes.…”

Section: Resultsmentioning

confidence: 99%

“…27,28 A reduced water content around protein molecules modifies the hydrogen bond interactions between water and protein, thereby increasing the denaturation temperature. 29 In addition, the application of vacuum provided a relatively inert atmosphere which may enhance the thermostability of enzymes. However, the activity of enzymes decreased quickly after 12 batches, and it reduced to 74% when reused in the 14th batch.…”

Section: Reusability Of Lipasementioning

confidence: 99%

High efficiency enzymatic synthesis of triglycerides with varying chain length and degree of unsaturation in a solvent‐free system under vacuum

Chen

Liu

et al. 2020

J of Chemical Tech & Biotech

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BACKGROUND Triglycerides with varying chain length and unsaturation degree are now in high demand in the food and pharmaceutical industries. Enzymatic catalyzed synthesis methods are favored over chemical‐based processes in the commercial production of triglycerides. This study found that the application of vacuum can improve the efficiency and productivity of enzymatic synthesis of various triglycerides using Novozym 435 (EC 3.1.1.3), an immobilized lipase B from Candida antarctica. The system was used for esterification of fatty acids varying in chain length, unsaturation degree, and oxidation degree. RESULTS The vacuum condition was not applicable to short chain fatty acids (C4–C6) due to loss of fatty acids in the reaction. The conversions of C4–C6 saturated fatty acids were low (15–35%) under atmospheric pressure. Under vacuum, high conversions (>95%) and triglyceride yields (approximately 90%) were achieved in the esterification of glycerol with several medium‐chain and long‐chain (C8–C18) saturated fatty acids. Moreover, unsaturation degree of fatty acids also affected the triglyceride yield. The triglyceride contents synthesized from C18:2n‐6 (76.17 ± 0.45%) was significantly lower (P < 0.05) than those from C18:0 (91.59 ± 1.58%) and C18:1n‐9 (90.77 ± 0.85%). Furthermore, fatty acids with a high oxidation degree had negative effect on triglyceride synthesis. This vacuum solvent free system allowed the synthesis enzyme to be reused 11 times without significant decrease in the lipase activity for esterification of fatty acids. CONCLUSION The studied vacuum facilitated triglyceride synthesis process provides an economically attractive yet sustainable solution for the production of a wide range of triglyceride products. © 2020 Society of Chemical Industry

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Drying in Biotechnology

Putranto¹,

Chen²

2019

Comprehensive Biotechnology

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Controlled water content for evaluation of denaturation temperature of freeze-dried enzymes

Cited by 7 publications

References 33 publications

Specific Interactions and Environment Flexibility Tune Protein Stability under Extreme Crowding

Specific Interactions and Environment Flexibility Tune Protein Stability under Extreme Crowding

High efficiency enzymatic synthesis of triglycerides with varying chain length and degree of unsaturation in a solvent‐free system under vacuum

Drying in Biotechnology

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