Evaluation method for catalytic hydrolysis performance of protease on chrome tanned collagen fiber

A technology for tanning collagen and catalyzing hydrolysis is applied in the field of catalytic hydrolysis performance evaluation and activity determination of chrome tanned collagen fibers by protease, which can solve the problem of lack of functional performance in the selection of wet blue leather enzyme softening technology and the blindness of control of softening conditions, etc. problems, to achieve the effect of flexible test methods and easy promotion and application

Active Publication Date: 2018-11-20
SICHUAN UNIV
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Problems solved by technology

[0005] The object of the present invention is to overcome the deficiencies in the prior art and provide a kind of protease to the catalytic hydrolysis performance evaluation of chrome-tanned collagen fiber and its application, to solve the problem of lack of quantitative characterization of protease on the performance of chrome-tanned collagen fiber The enzymatic softening technology of wet blue leather caused by the traditional method has the problem of blindness in the selection of protease and the control of softening conditions

Method used

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  • Evaluation method for catalytic hydrolysis performance of protease on chrome tanned collagen fiber
  • Evaluation method for catalytic hydrolysis performance of protease on chrome tanned collagen fiber
  • Evaluation method for catalytic hydrolysis performance of protease on chrome tanned collagen fiber

Examples

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Embodiment 1

[0033]Example 1——Catalytic hydrolysis performance evaluation and activity determination of different types of proteases on chrome-tanned collagen fibers

[0034] (1) Preparation of reagents

[0035] Prepare 0.1mol / L Braytan-Robinson buffer solution (B-R buffer solution): Weigh 6.183g boric acid and dissolve it in an appropriate amount of distilled water, add 11.529g phosphoric acid and 6.005g acetic acid, and dilute to 1L with distilled water, then use 4mol / L of NaOH solution to adjust the pH to 3.0, 7.0 or 9.0, respectively.

[0036] Prepare 0.08 g / mL NaCl solution: Weigh 0.5 g NaCl, dissolve it in distilled water and dilute to 100 mL.

[0037] Protease 537 was dissolved with pH 3.0 B-R buffer and diluted 100 times, and the supernatant was taken after centrifugation to obtain 537 enzyme solution; Protease AS1.398 was dissolved and diluted 100 times with pH 7.0 B-R buffer, and the supernatant was taken after centrifugation The supernatant was obtained as the AS1.398 enzyme ...

Embodiment 2

[0049] Example 2—Evaluation of catalytic hydrolysis performance and activity determination of alkaline protease 2709 on chrome-tanned collagen fibers under different pH conditions

[0050] The preparation of the chrome-tanned leather powder substrate, the determination of the concentration of hydroxyproline in the digestion solution and the calculation method of the enzyme activity are the same as those in Example 1, and the determination of the concentration of hydroxyproline uses the analysis method of an amino acid analyzer. The preparation of the reagents and the preparation of the reaction solution to be tested and the blank control solution are as follows:

[0051] (1) Preparation of reagents

[0052] Prepare 0.1mol / L Braytan-Robinson buffer solution (B-R buffer solution): Weigh 6.183g boric acid and dissolve it in an appropriate amount of distilled water, add 11.529g phosphoric acid and 6.005g acetic acid, and dilute to 1L with distilled water, then use 4mol / L of NaOH...

Embodiment 3

[0061] Example 3—Evaluation of catalytic hydrolysis performance and activity determination of alkaline protease 2709 on chrome-tanned collagen fibers under different temperature conditions

[0062] The preparation of the reagents, the preparation of the chrome-tanned leather powder substrate, the determination of the concentration of hydroxyproline in the digestion solution and the calculation method of the enzyme activity are the same as in Example 2, and the pH of the selected B-R buffer is 9.0. The steps for preparing the reaction solution to be tested and the blank control solution are as follows:

[0063] Weigh 5 parts of 100 mg chrome tanned leather powder as the substrate, add 5 mL of B-R buffer solution with pH 9.0, mix well, heat to 30, 35, 40, 45 and 50°C respectively, then add 1 mL of the Enzyme solution, in a water bath constant temperature oscillator under shaking conditions at 30, 35, 40, 45 and 50 ° C enzymolysis reaction for 4 hours, at this time the chrome tan...

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Abstract

The invention provides an evaluation and vitality measurement method for catalytic hydrolysis performance of protease on chrome tanned collagen fibers and applications The method includes the following steps: (1) a step of the preparation of a chrome tanned hide powder substrate, namely the step of adding a certain amount of a chrome tanning agent to perform tanning on hide powder, and performingfull washing, drying, crushing and filtering through 20-200 mesh screen; (2) a step of the preparation of a to-be-measured reaction solution, namely the step of adding 1-20 mL of a buffer solution into 50-200 mg of chrome tanned hide powder, uniformly mixing the mixture, heating the mixture to a measured temperature, adding 0.1-5.0 mL of a protease solution, performing oscillating reaction under the measured temperature, centrifuging after the reaction is finished, taking supernatant to perform digestion, and obtaining the to-be-measured reaction solution after the digestion solution is diluted; and (3) a step of hydroxyproline concentration determination and enzyme activity calculation in the digestion solution. The method is suitable for characterizing the hydrolysis activity of proteaseon chrome tanned collagen fibers and evaluating the effect of the protease on wet-blue leather in the leather field.

Description

technical field [0001] The present invention relates to a method for evaluating the catalytic hydrolysis performance of protease on chrome-tanned collagen fiber and measuring its activity and its application, in particular to a method for characterizing the hydrolysis activity of protease on chrome-tanned collagen fiber and the performance evaluation of protease on wet blue leather The method belongs to the field of measuring protease activity. Background technique [0002] With the increasing attention to environmental protection, more and more tanneries tend to purchase wet blue leather as raw materials for production. However, the purchased wet blue leather generally has insufficient dispersion of leather fiber structure, obvious defects such as blood tendons and acne scars, and the properties of wet blue leather from different sources are quite different due to different pretreatment processes. Defects such as mold spots, pigments, leather hardening, wrinkles and crease...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/68
Inventor 彭必雨张旭张茜张春晓李艳红
Owner SICHUAN UNIV
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