Storage stable liquid formulation comprising laccase
A liquid preparation, laccase technology, applied in the direction of enzyme, enzyme stabilization, oxidoreductase, etc., can solve the problems of short storage time and low level of residual enzyme activity
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Embodiment 5
[0044] The effect of increasing the pH from 9 to 10 is described in Example 5 and shown in Figure 4 middle. Myceliophthora thermophila laccase appeared to be more stable at pH 9.0, retaining 93% and 84% of the activity after storage at 25°C and 40°C, respectively, for 4 weeks.
[0045] Laccase preparation of polyporus sieve
[0046] In addition, as disclosed in WO 96 / 00290 (Novo Nordisk), the optimal pH of recombinant polyporus sieve laccase is 5 to 5.5 when using syringaldazine as substrate.
[0047] According to the present invention, the pH of the liquid formulation comprising laccase derived from Polyporus sieves and polyol should be more alkaline than pH 5 to 5.5, preferably about pH 5.5-11, especially about pH 6-9.5.
[0048] As shown in Example 1, the residual activity of Polyporus sieve laccase in 60% PEG6000 liquid at pH 9.0 was 100% after 14 days at 40°C, and 65% after 4 weeks.
[0049] Polyol
[0050] For improved storage stability of liquid formulations co...
Embodiment 1
[0228] Myceliophthora thermophila laccase and Polyporus sieveii laccase in buffer at optimal pH (without polyol) storage stability
[0229] The Myceliophthora thermophila and Polyporus sieves laccases were stored for 16 days at 40°C and their optimal pH (ie pH 6.5 and 5.5, respectively).
[0230] This Myceliophthora thermophila laccase was dissolved in 25 mM Tris / HCl buffer solution, the pH was adjusted to pH 6.5, and the activity was adjusted to 1000 LAMU / ml. Polypore sieve laccase was dissolved in 25 mM MES buffer, the pH was adjusted to pH 5.5, and the activity was 1000 LACU / ml.
[0231] The dissolved enzymes were placed at 4°C and 40°C, respectively, and samples were taken for analysis at the following time intervals: 24 hours, 3 days, 7 days and 16 days.
[0232] The relative residual activity refers to the corresponding formulation stored at 4°C (empty solution, the blind), ie the residual activity at 4°C is considered as 100% residual activity.
[023...
Embodiment 2
[0235] Storage Stability of Laccase from Polyporus sieves (14 days)
[0236] The storage stability of a liquid formulation comprising a high purity polyporus laccase (10 LACU / g) and two different polyols was tested at pH 7.0, 8.0 and 9.0.
[0237] The samples detailed in Tables 1 and 2 were prepared and stored for 14 days. Based on the laccase activity of the sample at 4°C under corresponding conditions, the remaining laccase activity after storage was calculated.
[0238] preparation
[0239] preparation
[0240] As can be seen from Tables 1 and 2, the storage stability is improved in the presence of polyols.
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