Optimizing the zymogram of exogenous proteases in broiler diets by in vitro simulated gastrointestinal digestion and response surface methodology

Abstract Low protein digestibility causes intestinal flora imbalance, wet litter, and nitrogen pollution. The present study aimed to explore the optimal zymogram of exogenous acid protease (ACP), neutral protease (NEP), alkaline protease (ALP), and keratinase (KEA) in corn-soybean based diets for br...

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Main Authors: Yang Liu, Shijie Liu, Fengming Chen, Shengli Liu, Qian Lin, Qiuzhong Dai
Format: Article
Language:English
Published: Nature Portfolio 2025-02-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-88954-w
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author Yang Liu
Shijie Liu
Fengming Chen
Shengli Liu
Qian Lin
Qiuzhong Dai
author_facet Yang Liu
Shijie Liu
Fengming Chen
Shengli Liu
Qian Lin
Qiuzhong Dai
author_sort Yang Liu
collection DOAJ
description Abstract Low protein digestibility causes intestinal flora imbalance, wet litter, and nitrogen pollution. The present study aimed to explore the optimal zymogram of exogenous acid protease (ACP), neutral protease (NEP), alkaline protease (ALP), and keratinase (KEA) in corn-soybean based diets for broilers. The hydrolysis performances of the four monocomponent proteases were presented by enzymatic hydrolysate gross energy (EHGE) and improved dry matter digestibility (IDMD), which were tested via the in vitro simulated digestion method. The optimal combination of the four proteases was predicted by the response surface method. Results showed that the optimal zymogram for 1 to 3-wk-old broiler diet was 1.80 U/g ACP, 2.30 U/g NEP, 29.30 U/g ALP, and 2.80 U/g KEA, and the EHGE and IDMD reached 94.65 Cal/g and 2.54%. The optimal zymogram for 4 to 6-wk-old broiler diet was 1.50 U/g ACP, 1.90 U/g NEP, 31.53 U/g ALP, and 3.10 U/g KEA, and the EHGE and IDMD reached 92.29 Cal/g and 2.47%. The performances of the predicted optimal zymogram were further verified by in vitro simulated digestion method. Collectively, the combined use of four proteases could improve the protein digestibility in broiler diets, which had better effect than monocomponent protease.
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spelling doaj-art-8abca6611d3a4d2d80df5fff4480e37b2025-08-20T03:04:01ZengNature PortfolioScientific Reports2045-23222025-02-0115111110.1038/s41598-025-88954-wOptimizing the zymogram of exogenous proteases in broiler diets by in vitro simulated gastrointestinal digestion and response surface methodologyYang Liu0Shijie Liu1Fengming Chen2Shengli Liu3Qian Lin4Qiuzhong Dai5Hunan Provincial Key Laboratory of the Traditional Chinese Medicine Agricultural Biogenomics, Changsha Medical UniversityDepartment of Chemical and Biological Engineering, University of SheffieldHunan Provincial Key Laboratory of the Traditional Chinese Medicine Agricultural Biogenomics, Changsha Medical UniversityShandong Lonct Enzymes Co. Ltd.Hunan Provincial Key Laboratory of the Traditional Chinese Medicine Agricultural Biogenomics, Changsha Medical UniversityDepartment of Animal Nutrition, Hunan Institute of Animal Husbandry and Veterinary MedicineAbstract Low protein digestibility causes intestinal flora imbalance, wet litter, and nitrogen pollution. The present study aimed to explore the optimal zymogram of exogenous acid protease (ACP), neutral protease (NEP), alkaline protease (ALP), and keratinase (KEA) in corn-soybean based diets for broilers. The hydrolysis performances of the four monocomponent proteases were presented by enzymatic hydrolysate gross energy (EHGE) and improved dry matter digestibility (IDMD), which were tested via the in vitro simulated digestion method. The optimal combination of the four proteases was predicted by the response surface method. Results showed that the optimal zymogram for 1 to 3-wk-old broiler diet was 1.80 U/g ACP, 2.30 U/g NEP, 29.30 U/g ALP, and 2.80 U/g KEA, and the EHGE and IDMD reached 94.65 Cal/g and 2.54%. The optimal zymogram for 4 to 6-wk-old broiler diet was 1.50 U/g ACP, 1.90 U/g NEP, 31.53 U/g ALP, and 3.10 U/g KEA, and the EHGE and IDMD reached 92.29 Cal/g and 2.47%. The performances of the predicted optimal zymogram were further verified by in vitro simulated digestion method. Collectively, the combined use of four proteases could improve the protein digestibility in broiler diets, which had better effect than monocomponent protease.https://doi.org/10.1038/s41598-025-88954-wIn vitro simulated digestionPoultry dietProteaseResponse surface methodologyZymogram
spellingShingle Yang Liu
Shijie Liu
Fengming Chen
Shengli Liu
Qian Lin
Qiuzhong Dai
Optimizing the zymogram of exogenous proteases in broiler diets by in vitro simulated gastrointestinal digestion and response surface methodology
Scientific Reports
In vitro simulated digestion
Poultry diet
Protease
Response surface methodology
Zymogram
title Optimizing the zymogram of exogenous proteases in broiler diets by in vitro simulated gastrointestinal digestion and response surface methodology
title_full Optimizing the zymogram of exogenous proteases in broiler diets by in vitro simulated gastrointestinal digestion and response surface methodology
title_fullStr Optimizing the zymogram of exogenous proteases in broiler diets by in vitro simulated gastrointestinal digestion and response surface methodology
title_full_unstemmed Optimizing the zymogram of exogenous proteases in broiler diets by in vitro simulated gastrointestinal digestion and response surface methodology
title_short Optimizing the zymogram of exogenous proteases in broiler diets by in vitro simulated gastrointestinal digestion and response surface methodology
title_sort optimizing the zymogram of exogenous proteases in broiler diets by in vitro simulated gastrointestinal digestion and response surface methodology
topic In vitro simulated digestion
Poultry diet
Protease
Response surface methodology
Zymogram
url https://doi.org/10.1038/s41598-025-88954-w
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