At present, the global real bromelain manufacturers are mainly located in Thailand and Indonesia. Chongqing Joywin Natural Products Co., Ltd. is one of the main manufacturers with crude enzyme facility located in Prachuapkhirikhan Thailand, too. In recent years, due to the increasing market demand of bromelain, there has been a shortage of supply. And large amount of Indian “Bromelain” entered the global market at very low prices. The pricing is at US$20~$60/kg for 2400GDU “Bromelain”.

After investigation, it was found that there are no factories producing bromelain in India. At the first beginning, the India “bromelain” activity test method was mainly based on enterprise standard, and these Indian companies cheated on the test method by adjusted conversion coefficient in the formula to achieve qualified enzyme activity detection.

 With more and more customers realized this is a wrong assay and found the real activity is only one-tenth of the true activity, the Indian manufacturers began blend papain with other enzymes as bromelain. As the GDU quantitative detection method of both papain and bromelain are the same, the Indian “bromelain” on the market is qualified with the enzyme activity. However as the papain produced in India has stink smell of protein fermentation, they usually use flavours and fragrances to cover the smell. This makes Indian “Bromelain” with both fermentation smell and flavour smell. While the true pure bromelain has the special frgrant smell of pineapple stem.

Identification of Bromelain

1. SDS-PAGE Analysis

Mark: M-protein marker，1-Bromelain（Joywin）（10μL）; 2-Bromelain（India manufacturer）（10μL）; 3-Bromelain（India）（10μL）; 4-Papain（10μL）

The molecular weight of Protein Marker bands from top to bottom in sequence is:

180 Kda, 130 Kda, 100 KDa, 70 KDa, 55KDa, 40 KDa, 35 KDa, 25 KDa, 15 KDa, and 10 KD.

The SDS-PAGE Analysis results shows that the characteristic band of bromelain is at about 25Kda, there are 3 main characteristic bands of bromelain, while papain has multiple bands from 13~30.

2. MALDI-TOF MS Identification

From the MALDI-TOF MS chromatogram could see, bromelain only has one characteric peak, at about 24.5Kda, while papain has three characteric peaks, at around 13.4Kda, 23.7Kda and 26.7Kda. It further indicates that the essence of Indian "bromelain" is papain.