Characterisation of the lipolytic enzymatic activities of fungal rizoenzymes from Rhizopus oryzae in comparison to pancreatin from pigs

C. Schön, R. Wacker, M. Rothe, B. Lipowicz, A. Iphöfer
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Abstract

In case of exocrine pancreatic insufficiency (EPI), the replacement of digestive enzymes with, for example porcine pancreatin or fungal rizoenzymes, is unavoidable under certain conditions. Current guidelines indicate that preparations from porcine pancreas have more advantageous physicochemical properties compared to those from fungi, especially at high bile salt concentrations, and that the latter can, therefore, only be used clinically to a limited extent. Since rizoenzymes are increasingly used in clinical practice, the present in vitro study investigated efficiency of enzymatic activity of rizoenzymes in comparison to pancreatin under various physiological and partly extreme environmental conditions. The lipolytic properties of two typical preparations containing digestive enzymes from porcine pancreatin and rizoenzymes were compared (same dosage of activity units) at different pH values (pH 3–9), with the addition of different bile salt concentrations (0–15 mmol/L sodium taurocholate). Lipolytic activity was measured by quantifying the fatty acids released from olive oil substrate by liquid chromatography–tandem mass spectrometry after enzymatic digestion. For both enzyme preparations, the lipolytic activity maximum was reached at pH 7 with comparable fatty acid conversion rates (57% pancreatin, 58% rizoenzymes). However, in contrast to pancreatin, rizoenzymes were already active for certain fatty acids from pH 3 to 4. At a bile salt concentration up to 10 mmol/L taurocholate, there was an increase in activity of both enzyme preparations (rizoenzymes 69% vs. pancreatin 58% enzymatic conversion). Only at rather unphysiological concentration of 15 mmol/L during EPI, there was a slight decrease in activity (to 56%) for the rizoenzymes. Rizoenzymes are an alternative therapeutic option for EPI. Relevant differences between rizoenzymes and pancreatin with regard to lipolytic activity under different physicochemical conditions could not be demonstrated in this study, whereas a potential advantage in favor of rizoenzymes activity at already low pH values was observed.
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来自根瘤蚜的真菌涟酶的脂肪分解酶活性特征与猪胰蛋白酶的比较
在胰腺外分泌功能不全(EPI)的情况下,在某些条件下不可避免地要用猪胰蛋白酶或真菌酵素等消化酶来替代。目前的指导方针表明,猪胰腺制剂的理化性质比真菌制剂更有优势,尤其是在胆盐浓度较高的情况下,因此后者只能在有限的范围内用于临床。由于胰酶在临床上的应用越来越广泛,本体外研究调查了胰酶与胰蛋白酶在各种生理条件和部分极端环境条件下的酶活性效率。 在不同的 pH 值(pH 3-9)下,加入不同浓度的胆盐(0-15 mmol/L 牛胆酸钠),比较了两种含有猪胰蛋白酶和涟漪酶消化酶的典型制剂(活性单位剂量相同)的脂肪分解特性。通过液相色谱-串联质谱法对酶解后橄榄油底物释放的脂肪酸进行定量,从而测定脂肪分解活性。 两种酶制剂的脂肪分解活性在 pH 值为 7 时达到最大值,脂肪酸转化率相当(57% 的胰蛋白酶和 58% 的涟漪酶)。当胆汁盐浓度达到 10 mmol/L 牛磺胆酸盐时,两种酶制剂的活性都有所提高(胰蛋白酶的酶转化率为 69%,而胰酶的酶转化率为 58%)。在 EPI 期间,只有在 15 毫摩尔/升的非生理浓度下,胰酶的活性才略有下降(降至 56%)。 芮佐酶是 EPI 的另一种治疗选择。本研究未能证明在不同的理化条件下,胰酶和胰蛋白酶在脂肪分解活性方面存在相关差异,但观察到在 pH 值已经很低的情况下,胰酶的潜在优势有利于胰蛋白酶的活性。
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来源期刊
European Pharmaceutical Journal
European Pharmaceutical Journal Pharmacology, Toxicology and Pharmaceutics-Pharmacology, Toxicology and Pharmaceutics (all)
CiteScore
0.60
自引率
0.00%
发文量
16
期刊介绍: European Pharmaceutical Journal publishes only original articles not previously published and articles that are not being considered or have not been submitted for publication elsewhere. If parts of the results have been published as conference abstract or elsewhere, it should be stated in references. The ethical standards of the Helsinki-Tokio Declaration should be kept. This should be mentioned in the Methods of manuscript. Reviews are published only on request. Authors, whose submitted research work was performed with the support of a company, should indicate this in Conflict of Interest.
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