Bromelain powder, also known as pineapple enzyme or bromelain enzyme, is a natural plant protease extracted from the stems, leaves, and skins of pineapples. The molecular weight of bromelain is about 33,000. It has an isoelectric point of 9.55. The best quality bromelain is processed from the middle stems of pineapples and is concentrated by ultrafiltration and freeze-dried at low temperatures. Bromelain appears as a light gray powder with a slightly peculiar odor. The optimal pH values for bromelain's action on casein, hemoglobin, and BAEE are between 6 and 8. For gelatin, the optimal pH value is 5.0. Enzyme activity is inhibited by heavy metals. It is slightly soluble in water but insoluble in alcohol, acetone, chloroform, or ether. Bromelain is a glycoprotein.
Bromelain is composed of an enzyme system consisting of multiple enzymes with different molecular weights and structures. It contains at least five proteolytic enzymes. In addition to these, it may also contain phosphatase, peroxidase, cellulase, other glycosidases, and non-protein substances.
Bromelain has been widely used in the food and pharmaceutical industries. In the food industry, it can be used for beer clarification, digestive aid in medicine, and anti-inflammatory and detumescence.
How to identify real Bromelain powder?
Identifying bromelain usually requires professional laboratory testing, including but not limited to the following steps:
1. Activity detection: The authenticity of bromelain is evaluated by measuring the activity of the enzyme. Bromelain has a specific enzyme activity, which can be detected by standard enzyme activity determination methods.
2. Purification effect comparison: The authenticity of bromelain can be identified by comparing the purification effects of bromelain obtained by different purification methods. For example, the purification effects of different methods such as kaolin adsorption method, ultrafiltration concentration organic solvent precipitation method and nano-titanium dioxide adsorption method will be different.
3. Stability study: The stability of bromelain under specific conditions can be used as an indicator to identify its authenticity. For example, the enzyme activity of bromelain is relatively stable at pH 5.0-6.5, and the enzyme activity stability is best at pH 6.0.
4. Chemical analysis: The content of specific components in bromelain can be detected by chemical analysis methods, such as high performance liquid chromatography (HPLC), so as to identify its authenticity.
5. Bioassay: The biological activity of bromelain can be detected by bioassay methods, such as enzyme-linked immunosorbent assay (ELISA), so as to identify its authenticity.
6. Regulatory agency certification: Checking whether the product has regulatory agency certification, such as the approval number of the Food and Drug Administration, is also an important way to identify authenticity.
It should be noted that these methods usually require professional equipment and technicians to carry out, and ordinary consumers may find it difficult to complete them on their own. Therefore, when purchasing, it is a relatively simple identification method to choose reputable suppliers and brands and check the product's certification and test reports.
Why is the Bromelain powder in India so cheap? Is it real?
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.
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Type |
India "Bromelain" manufacturer Assay |
International Common Assay |
Memo |
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Assay comes from |
Indian manufacturer's GDU Assay |
International Common GDU Assay |
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Similarities |
Substrate |
Gelatin |
Gelatin |
Use same substrate |
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Definition of enzyme activity |
Unit Definition (Gelatin Digestion Unit-GDU): One unit will hydrolyze 1.0mg of amino nitrogen from gelatin in 20 minutes at pH 4.5 at 45℃. |
Unit Definition (Gelatin Digestion Unit-GDU): One unit will hydrolyze 1.0mg of amino nitrogen from gelatin in 20 minutes at pH 4.5 at 45℃. |
GDU enzyme activity definition same |
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Test Steps |
Use NaOH solution for titrant test |
Use NaOH solution for titrant test |
Main test method are the same |
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Differences |
Titrant concentration |
0.05mol/L NaOH Titrant |
0.1mol/L NaOH Titrant |
The concentration of NaOH titrant solution of common GDU assay is double than the Indian GDU assay. |
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End point of titrant |
End point PH 7.8 |
End point PH 9.0 |
Different pH value on end point identification |
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Formula |
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Different computational formula |
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Indian Assay: No explainations to the computational formula Based on the test procedure and the definition of enzyme activity we assume： T= Test titer (ml 0.05 N NaOH) B= Blank titer (ml 0.05 N NaOH) ml 0.1- Initial weight of enzyme (g) |
T= Test titer (ml 0.1 N NaOH) ml B= Blank titer (ml 0.1 N NaOH) ml N=Normality of standardized NaOH (mol/L) M=Initial weight of enzyme (g) 14-Molecular weight of nitrogen; 50-Dilution ratio of the enzyme sample |
According to Indian manufacturer's Assay, and combined with international common GDU enzyme activity definition, the digit 700 in the formula should be(14×0.05×100=70), but they mark it as 700. This leads to a wrong result which is 10 times increased. |
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 powder
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.
Characteristics of Bromelain
(1)Unique mechanism of action
Bromelain has a unique action principle. It can hydrolyze polypeptides into low molecular weight peptides. This characteristic reflects its strong proteolytic ability. It can also hydrolyze amide bonds and esters. This multi-functional hydrolysis ability makes bromelain stand out in the field of enzymes. For example, in the food industry, it can be used for beer clarification by breaking down proteins that cause turbidity, making the beer clearer and more stable. In the medical field, it can help with digestion by breaking down proteins in food into smaller peptides that are easier for the body to absorb.
(2)Affected by multiple factors
Bromelain is affected by various factors. The pH value has a significant impact on its activity. The optimal pH for bromelain's action on different substrates varies. Generally, the optimal pH range is between 6 and 8 for casein, hemoglobin, and BAEE, while for gelatin, the optimal pH is 5.0. Temperature also plays a crucial role. The optimal reaction temperature is 55°C. At higher or lower temperatures, its activity may decrease. For instance, if the temperature is too high, the enzyme may denature and lose its activity. Metal ions can also affect bromelain. Mg²⁺ and Ca²⁺ have inhibitory effects at high concentrations but can promote activity at low concentrations. For example, when the concentration of Ca²⁺ is 2mmol/L, it has the most obvious promoting effect. On the other hand, heavy metals can inhibit the enzyme activity. EDTA can chelate the metal ions required for bromelain reaction, thereby reducing its activity. In conclusion, understanding these factors that affect bromelain can help us better use and control this enzyme.
Application of Bromelain powder
Bromelain has extensive applications in various fields, especially in the food processing industry. In food processing, bromelain can be used for tenderizing meat. By breaking down the proteins in meat, it makes the texture softer and more palatable. For example, in the production of some processed meat products, adding a certain amount of bromelain can significantly improve the quality of the meat.
It can also be used in the brewing industry. Bromelain can help clarify beer by removing proteins that cause turbidity, resulting in a clearer and more stable product. According to research, the application of bromelain in beer clarification can improve the visual quality of beer and extend its shelf life.
In addition to the food industry, bromelain also has applications in the pharmaceutical field. It has anti-inflammatory and detumescence effects. In some cases, it can be used as an adjunct treatment for certain inflammatory diseases. For example, in the treatment of sports injuries or post-operative inflammation, bromelain may be used to reduce swelling and pain. Moreover, bromelain may have potential applications in other areas. For instance, in the cosmetic industry, it may be used in some skincare products due to its ability to promote skin metabolism and improve skin texture.
In conclusion, bromelain's unique properties and multiple functions make it an important enzyme with wide-ranging applications in different industries.
JOYWIN founded in 2013 is an innovation-driven biotechnology company. The JOYWIN Bromelain factory located in Thailand makes use of local abundant resources to provide customers with various specifications of bromelain products. From 200GDU/g to 2400GDU/g. Holding bromelain workshops, plant protease workshops, and warehouses also own cutting-edge facilities and strict quality control systems. As one of four global bromelain manufacturers, we are FSSC22000, ISO9001, ISO14001, ISO22000, BRC, and Cgmp-certified factory. If you want to know more about Bromelain Powder or are interested in purchasing it, you can email at contact@joywinworld.com. We will reply to you as soon as possible after we see the message.