College of Chemistry and Chemical Engineering, Lingnan Normal University, Zhanjiang, China.
Department of Industrial Biotechnology, Prince of Songkla University, Hat Yai, Thailand.
J Food Biochem. 2019 Nov;43(11):e13011. doi: 10.1111/jfbc.13011. Epub 2019 Aug 8.
Bromelain is a type of protease found in both fruits and stems of pineapples. Stem bromelain has been extensively studied and is commercially available for applications in various industries. In contrast, studies of fruit bromelain are quite limited since most of pineapples have been consumed freshly, canned or juiced. Nowadays, the consumption of canned fruits, including canned pineapples has decreased greatly. Fruit bromelain could be a new growth point for pineapple industry. In this study, fruit bromelain was extracted from the pineapple juice of Phuket variety and some of its properties were studied. The enzyme was purified by precipitation using ammonium sulfate fractionation followed by ion-exchange and gel filtration chromatography. Consequently, the protease purification level was increased by 95.2 fold. The final specific activity was getting to 448,590 U/mg on average, dominated by cysteine proteases, with optimal activity at 45°C and pH ranging from 6 to 8. The study facilitates the molecular and application research of fruit bromelain. PRACTICAL APPLICATIONS: The research has been carried out at Funong Food Technology Co., Ltd., Guangdong, China, which produces primarily pineapple chunks and juice. As plenty of by-products, like peels and cores of pineapples, are produced, the techniques are employed to extract bromelain from the by-products. The techniques reported in this work are not new or advanced, however, they are applicable during the manufacturing process and the related equipment is easy to set up and operate. What's more, the practical application of the techniques is cost-effective for the manufactories. Take Funong Food Technology Co., Ltd. as an example, they was using 80% saturation ammonium sulfate to precipitate protein from pineapple juice and obtained a bromelain with activity of approximately 8,000 U/mg and yield of 1.7 kg per ton of juice. With the application of the techniques reported in this work, bromelain was first extracted by ammonium sulfate gradient precipitation, and then purified through ion-exchange and gel filtration chromatography. Each step of precipitation and purification generates a different level of activity and recovery of bromelain, ranging from around 2,506 to 448,590 U/mg, which allows for the production of bromelain according to the requirement of the market and brings more profits.
菠萝蛋白酶是一种存在于菠萝果实和茎中的蛋白酶。茎菠萝蛋白酶已被广泛研究，并可在各个行业的应用中商业化。相比之下，由于大多数菠萝已被新鲜食用、罐装或榨汁，因此对果实菠萝蛋白酶的研究相当有限。如今，包括罐装菠萝在内的罐装水果的消费大大减少。水果菠萝蛋白酶可能成为菠萝产业的新增长点。在这项研究中，从普吉岛品种的菠萝汁中提取了果实菠萝蛋白酶，并对其一些性质进行了研究。该酶通过使用硫酸铵分级沉淀进行纯化，然后通过离子交换和凝胶过滤色谱进一步纯化。因此，蛋白酶的纯化水平提高了 95.2 倍。最终平均比活达到 448,590 U/mg，主要为半胱氨酸蛋白酶，最适活性在 45°C 和 pH 值为 6 到 8 之间。这项研究为果实菠萝蛋白酶的分子和应用研究提供了便利。
本研究由广东丰农食品科技有限公司进行，该公司主要生产菠萝块和菠萝汁。由于产生了大量的副产品，如菠萝皮和菠萝心，因此采用这些技术从副产品中提取菠萝蛋白酶。本工作中报道的技术并不新颖或先进，然而，它们适用于制造过程，相关设备易于设置和操作。更重要的是，这些技术的实际应用对制造厂来说具有成本效益。以丰农食品科技有限公司为例，他们使用 80%饱和度的硫酸铵从菠萝汁中沉淀蛋白质，获得了一种活性约为 8,000 U/mg 的菠萝蛋白酶，每吨果汁的产率为 1.7 公斤。应用本工作中报道的技术，首先通过硫酸铵梯度沉淀提取菠萝蛋白酶，然后通过离子交换和凝胶过滤色谱进行纯化。沉淀和纯化的每一步都会产生不同水平的菠萝蛋白酶活性和回收率，范围从 2,506 到 448,590 U/mg，这使得可以根据市场需求生产菠萝蛋白酶，并带来更多的利润。
