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THE EXTRACTION OF PINEAPPLE (ANANAS COMOSUS) FRUIT JUICE WITH HIGH CONCENTRATION OF BROMELAIN ENZYME

Simposium Nasional Kimia Bahan Alam XIX(SimNasKBA-2011)

Samarinda, 11-12 Oktober 2011

Presenter: Dr.rer.nat.Maruli Pandjaitan

Department of Food Technology, Faculty of Life SciencesSwiss German University

S W I S S G E R M A N U N I V E R S I T Y

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OVERVIEW

• Background

• Research Purpose

• Research Problem

• Methodology

• Result & Discussion

• Conclusion

• Recommendation


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BACKGROUND

Pineapple fruit

Indonesia as a tropical country

Bromelain enzyme


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BACKGROUND

• Enzyme is protein that catalyze chemical reaction• Bromelain is a protease enzyme• Protease breaks down protein to its simpler

structure• Hence, a good extraction process could

obtain pineapple juice extract with high bromelain enzyme activity• Main challenge is that bromelain enzyme is

highly heat sensitive: exposure to high temperature will destroy the enzyme


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BACKGROUND

Has been used in various traditional medicine

• Anti-inflammatory agents

• Digestive aid

• Anti tumor agents

• Antiviral agents

However it also exists in fruit

Bromelain mostly in stem parts of the fruit


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BACKGROUND: HIV virus that causes AIDS

• Protease enzyme cleaves protein• Virus receptor is made of

protein• The receptor is important

to the life cycle of the HIV virus when it attacks the human immune system (CD4+)• Protease damage the

virus


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BACKGROUND: HIV virus that causes AIDS

CD4+ Before Pineapple Juice Diet

CD4+ After Pineapple Juice Diet

135 sel/uL 647 sel/uL (4 months)

<200 sel/uL 361 sel/uL (6 week)

<200 sel/uL 723 sel/uL (4 months)





• CD4+ changes occur in HIV patients after consumption of pineapple juice

• Normal: 410-1500 cell/μL

Source: Maruli Pandjaitan, Tutun Nugraha, Kezia Pamudja (2011)


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RESEARCH PURPOSE

To produce an instant drink product in the form of powder from pineapple fruit with

high bromelain enzyme activity

Advantages: • Readily available• Long storage time• Can be used for

medicinal purposes


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RESEARCH PROBLEM

1. Determination of fruit juice extraction & filtration that retains/protects enzyme activity

2. Retainment of bromelain enzyme activity in the dried product because bromelain is heat unstable

3. Fruit juice drying problem Stickiness


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METHODOLOGY

EXTRACTION

Juice Extraction

Juice Filtration

Drying:- Spray Drying- Freeze Drying

ANALYSISTotal Dissolved Solid

Protein Content

Enzyme Volume Activity

Moisture Content of powder

Solubility of instant powder


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METHODOLOGY

Additives in Spray Drying• Maltodextrin (MD) (5%, 15%, 25%, 35%)Prevent stickiness • ZnSO4 (5%, 10%, 20%, 40%, 80%)Protect the enzyme

Additives in Freeze DryingMaltodextrin (MD) (0%, 15%, 25%, 35%)Prevent stickiness

BÜCHI 190 Mini Spray Dryer

Freeze Dryer Telstar Lyo Alfa 15

T in: ±190oCT out: ±80oCRate: 1L/hour

T: -40oCP: 5 barTime: 4 days


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JUICE EXTRACTION

+ aquadest 1:1 ratio

Fresh Juice

Fresh Fruit Peeling “Eye” Removal Cutting


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JUICE FILTRATION

Fresh Juice

Filtered Juice

Filter 36 & 20 μm

Solid Waste 1

+ aquadest1:0.5 ratio

Blend &Filter (36 & 20 μm)

Solid Waste 2

+ aquadest1:0.5 ratio

Blend &Filter (36 & 20 μm)

Solid waste still retains enzyme activity so it should be re-processed


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RESULTS & DISCUSSION

Enzyme Volume Activity (U/ml):Enzyme activity per volume of a sample

Enzyme Specific Activity (U/mg protein):Enzyme activity per protein weight of a sample

• Filtered Juice is chosen as the best sample

• Fiber might clog the nozzle in spray drying

• Low protein in waste but the activity is already very low


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SPRAY DRYING with Maltodextrin (MD) variation

MD 5%20.7 gr

MD 15%34.1 gr

MD 25%48.7 gr

MD 35%59.8 gr

• 0% MD unsuccessful (sticky)• MD >15% has better

powder characteristics• Smooth powder,

forms no agglomeration for MD > 15%• All has good

solubility during reconstitution in water


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SPRAY DRYING with salt variation

ZnSO4 5%57.2 gr

ZnSO4 10%70.3 gr

ZnSO4 20%91.8 gr

ZnSO4 40%127.6 gr

ZnSO4 80%198.2 gr

• Protect enzyme during spray drying• Improve stability

of the enzyme in storage time• No stickiness

formed in all the samples


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SPRAY DRYING: Volume & Specific Activity

Best powder

• Activity decreased as more MD added• Smooth

powder from MD 15% onward


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SPRAY DRYING: Volume & Specific Activity

• Salt as activator• Low

sensory properties (Unsuitable taste by consumer)


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FREEZE DRYING

MD 0%3.8 gr(Unsuccessful)

MD 15%17.3 gr(Unsuccessful)

MD 25%26.9 gr

MD 35%35.5 gr

• MD 0% & 15% are not in powder form• Insufficient

maltodextrin• MD 25% & MD

35% has good powder texture


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FREEZE DRYING: Volume & Specific Activity

• No significant difference in volume activity• MD 25%

has better activity


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COMPARISON of Moisture Content Analysis

Sample Moisture Content (%)

Spray Drying(30 minutes process,

each 500 ml)

Maltodextrin 5% 5.9 ± 0.3




Maltodextrin 15% + Salt 5% 9.2 ± 0.1





Freeze Drying(48 hours process,

each 100 ml)




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COMPARISON of Solubility Analysis

Sample Solubility (gr/ml)

Spray Drying










Freeze DryingMaltodextrin 25% 0.45 ± 0.07

Maltodextrin 35% 0.37± 0.05


Volume Activity % Protection Comparison for Spray Drying & Freeze Drying Process

0%

10%

20%

30%

40%

50%

60%

70%

80%

90%

100%

MD 5% MD 15% MD 25% MD 35% MD 25% MD 35%

Spray Drying Freeze Drying

Types of Sample

Prot

ectio

n (%)

COMPARISON of Enzme Volume & Specific Activity

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• Spray dried with 15% MD is the best

• Protecting enzyme

• Good powder characteristics

• Shorter time of drying process


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CONCLUSION

• In this work, pineapple juice instant powder has been produced with spray dry & freeze dry process

• Stickiness of fruit juice during spray drying could be avoided by using maltodextrin.

• Maltodextrin & salt (ZnSO4) could protect enzyme, however due to its low sensory property, maltodextrin was preferred.

• Spray drying process with 15% maltodextrin was found to be the best drying process because it was able to protect the enzyme activity with good powder characteristics & short drying time.


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RECOMMENDATION

• Use of other types of maltodextrin (different DE value),• Analysis of other nutrient like Vitamin C or antioxidant • Use of other drying method like spray chill.• Analysis of the stability of the dried product over time as

well as the choices of packaging type that would best protect the product.

• The study should also investigate acceptability by potential user through sensory tests.

• Addition of sugar to improve acceptability of user, its impact on the activity of enzyme, however, should be studied.


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