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Omar, R., Ali Rahman, Z., Latif, M.T., Lihan, T. and Adam J.H. (Eds.): 244-250

Proceedings of the Regional Symposium on Environment and Natural Resources (Vol. 1)

10-11th April 2002, Hotel Renaissance Kuala Lumpur, Malaysia.

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COMMERCIALISATION OF PAPAIN ENZYME FROM PAPAYA

Krishnaiah D., Awang B., Rosalam S. and Buhri A.

Chemical Engineering Programme

School of Engineering and Information TechnologyUniversiti Malaysia Sabah

88999 Kota KinabaluE-mail: [email protected]

ABSTRACT

The potential application of papain enzyme from papaya is explored in this study forcommercialisation. The papaya varieties suitable for Sabah especially are broughtabout in the production application. The various parameters of spray dryer are

critically discussed for obtaining the papain dry powder, characteristics of sizedistribution and activity are mentioned in view of the production growth and theproduct demand in the world market. With its popularity and price in the market,

papain can be a potential economic enzyme in the world.

INTRODUCTION

Sabah state has considerable potential of papaya resource, a sustainable and assured

tropical product in this region. Perspectives for its commercial utilisation, forms a value

added product and is discussed in this paper from technical point of view. The green

papaya fruits are source for papain enzyme, which are valued as an industrial product inpreparing various digestive enzymes in pharmaceuticals, foods, chill proofing of beer and

in meat tenderising Arnon (1970). Besides, it is also used in textiles, detergents,

adhesives manufacture, dental, face cream, toothpaste and sewage disposal. It is also

grown in Johore and parts of Perak and Selangor.

Production of papain requires considerable technical parameters starting from the plant.

The latex is collected from the papaya green fruit after 2-3 months fruit is set. Different

methods are used in lancing the fruit Ram (1996). The collected latex is dried and treated

with suitable solvents in order to validate the activity of papain enzyme to be used in the

different applications. Yield of papin depends on cultivator, time of taping, nutritional

status of plant and the region. It is reported that there is positive correlation between thefruit size, papain yield, rainfall in respect of papain enzyme yield.

Papaya is also good source of pectin. Lanced fruits after papain extraction can be utilised

for pectin extraction. There are several uses of pectin. Commercially it is used in the food

preparation and as emulsifying agent. Mukherjee and Kiewitt (1996) reported on the

specificity of papaya latex as biocatalyst in the esterfication of several fatty acids with n-

butanol.

Up on the critical literature survey, the details on the enzyme collection, purification and

activity are not available. Most of the data published only pertaining of the general topic

of interest with protocols. Several patents reported in the literature from different
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countries as an intellectual property. In view of the technical commercial importance, the

research work is under taken to carry out a through investigation and to obtain a reliable

scientific and technical data commercialisation, as there is good potential for revenue in

the over seas market.

CULTIVARS

Papayas have exacting climate requirements for vigorous growth and fruit production.

They must have warmth throughout the year. Cold and wet soil is almost lethal. Cool

temperature will also alter fruit flavour. Papaya makes excellent container and

greenhouse specimens where soil moisture and temperature can be moderated. The yield

of papaya in the first year varies between 15-20 tonnes per hectare and in the second year

30-50 tonnes per hectare and in the third year 10-30 tonnes per hectare.

There are several cultivars of papaya, both local and inter continental introduced inMalaysia. These introduced cultivars include Cibinong and Paris, from Indonesia; sunrise

solo, Higgins and Wilde from Hawaii; Sunnybank from Queensland; Honewdew from

India; Maradol from Cuba and Kokdum from Thailand. Higgins and Wilder are derived

from the small-fruited solo parents. The local cultivars include Subang, Sitiawan and

Taiping.

Subang: This is short tree start bearing fruit at about 9-10 months old, at about 50-50 cm

from the ground. The fruit is long and weights about 1 kg, the flesh is yellowish, firm but

not very sweet.

Sitiawan: It is medium height, starts bearing fruit about 10-11 months old, at about 80 cm

from the ground. The fruit is long and weights about 2 kg. The flesh is red and firm but

rather flat in taste. It is suitable for processing.

Exotica: A pure line variety developed from a backcross programme involving the

Hawaiian Sunrise Solo and Subang 6. The plant is of medium height and starts flowering

after 4 months, fruits are harvested after 9 months from planting. The fruit weights about

600 to 800 gm with arangey red flesh and high sugar content.

Vista Solo: Medium to large fruit depending on climate, 5 inches wide, up to 18 inches

long. Skin yellow, flesh orange to yellow-orange. Hardy, compact Solo type producinghigh quality fruit. Need fairly hot weather to develop sweetness. Self-fertile. Originated

in Vista,Calif. By Ralph Corwin.

EXTRACTION OF PAPAIN

The papaya fruit is very rich in papain. The greener the fruit the more active is the papin.

This protein digestion enzyme is found in the latex and is very abundant in green, unripe

fruits. The extraction of papaya enzyme was carried out by collecting the latex from

green papaya from a suitable size and age having 2 to 3 months after the fruit is set..

Different methods are employed in getting the latex. A suitable method established by


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using the fine razor blade or with a stainless steel knife for lancing the fruit. Four

longitudinal skin deep incision on fruit from the end to the fruit tip are given. Such

incision is repeated on 3-4 subsequent occasions at an interval of 4-5 days. A plastic

container is useful for collecting the latex.

The collected latex is dried in the vacuum oven in the range of 65C-80C. The enzyme

is extremely temperature stable in comparison to other proteases. Effective activity is

demonstrated over the temperature range of 10C-90C at 6-7 pH. After thoroughly

drying the latex becomes tan to light brown colour. This product is stable at 5C for 6-12

months. To enhance the stability as well as solubility it may be advantageous to convert

the crystalline papain to its mercury derivative. The temperature above 90C rapidly

inactivate the enzyme. The papain is purified by affinity chromatography Blumberg

(1970).

COMMERCIAL PRODUCTION : SPRAY DRYER DESIGN CONSIDERATIONS

Spray drying has achieved wide acceptance for drying the papain latex. The feed is

reduced to a fine spray ,mixed with a hot air stream, dried and separated as a dry product.

The spray dried papain has high solubility and reaction rates due to its uniformly sized

and fine particles. Spray dryers are especially adapted for large production capacity.

Spray drying produces fine particles by atomizing the papain latex and evaporating

moisture from the resulting droplets by suspending them in a hot gas. The production of

dry, spherical particles from a latex feed in a single processing step makes spray drying a

unique and important unit operation. Figure 1 is a schematic of a typical spray dryer, anexperimental setup at our laboratory .

Figure1: Experimental set up- spray dryer


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The liquid feed is pumped to an atomizer, which breaks up the feed into a fine spray and

ejects it into the first spray chamber, where it is mixed with hot drying air. Moisture is

evaporated from the spray and the droplets are transformed into dry particles. The

separation of the particles from the drying air and their subsequent collection will either

take place from the main chamber in the case of coarser particles and the fine particlesfrom the cyclone separator.

A key characteristic of the process is that atomization produces spherical droplets that dry

to form spherical particles. Final particle size may be controlled to some extent by

controlling the drop size produced by atomization in the range of 10500 microns. If

required, the spray dryer can be designed to produce large agglomerates, which are highly

desirable for some applications. Because of the high feed moisture content compared with

other drying and de watering processes, the energy requirements of spray drying are

relatively high. Within a spray chamber, droplets have an in-flight residence time ranging

from about five seconds to 50 seconds. This short-residence-time drying is good for

papain a heat-sensitive material but poor for the removal of tightly bound moisture. Toachieve low moisture contents, it may be necessary to follow the spray dryer by a dryer

with a long residence time. Although most spray drying operations involve the drying of

water-wetted material in hot air, materials wetted with other solvents can be dried in the

same way. The distribution of particle size in the spray dryer is shown in Figure 2.

Figure 2: Typical particle size distributions in the spray-dried product.

Ultrasonic technique can improve the spray drying process. Drying of papain enzyme is

greatly enhanced by using high power air borne ultra sound Ensminger (1988). The heat

transfer from the air simply pushes much of the moisture content out. On the other hand,

the ultra sound waves increase the diffusion rate and also generate micro streaming of air

in the vicinity of wet particles. Thus the time of drying is reduced by a factor of 2-3.


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CHARACTERISTICS

This enzyme has the property of substantial breaking the molecules. The use of itsproperty is estimated in the commercial application essentially in the soap and detergent

powder manufacture. Present market products are made mostly by raw papain. The

effectiveness of the enzyme can be improved by modifying and activating. Papain has

wide specificity. Arnon (1970) has indicated that it will degrade most protein substrates

more extensively than pancreas protease. Papain is activated by cystein,sulfide and

sulfite. It is enhanced when heavy metal binding agents such as EDTA are also present.

Kirschenbau (1971) indicated that N-bromosuccinimide enhances the activity.

Substances which react with sulfhydryl groups including heavy metals,carbonyl reagents

reported by Morihara (1967). It has been mentioned by Westerik and Wolfenden (1972)

that aldehydes exhibited as papain inhibitors and Wijdenes (1973) reported onbenzoylamidoacetonitrile as an inhibitors. These inhibitors have considerable effect

when the papain is used in the pharmaceutical applications. The general characteristics of

the two derived enzymes vary to some extent since they all have different temperatures of

inactivation and operate with different kinetics when applied.

MARKET OVERVIEW

Enzyme is in great demand in the international market essentially U.K.,U.S.A., Europe

and Gulf Countries. The present exporting counties are Zaria, Tanzania, Kenya , Israel,

Philllippines, Srilanka, India and Cameroon earning handsome foreign exchange. With

the available papaya in Sabah and other parts of Malaysia, it is highly profitable to

manufacture papain where the fruit production is in abundance possible as it is a tropical

region.

In Malaysia the production of fruits saw an increase of 6.2 percent annually from one

million tones in 1996 to 1.4 million tones in 2000 while harvested area grew by 4 percent

per annum from 244,500 hectares in 1996 to 297,400 hectares in 2000. A total of 15 fruit

types were promoted for commercial cultivation including papaya.

The food and pharmaceutical enzyme market shows a good expansion potential in thefuture. The enzyme market is growing at 3% to5% annually. The market potential in the

Europe and U.S.A is in great demand. Preliminary analysis of income for the cultivator

shows that in one acre of papaya plantation, an amount RM 40,000 revenue could be

obtained.

In 1992, a total quantity of 93,930 tonnes of papaya were produced in Malaysia and

23,225 tonnes of papaya were exported to foreign countries Singapore, Hong Kong and

Western Europe. The following Figure3 shows the Malaysia production and projections

in the near future in million MT.


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CONCLUSIONS

The study investigated the production and technical aspects of papain enzyme from

papaya enables the importance of the product in the commercialisation. The spary dryer isused in the large scale commercial producrtion. It is evident from the research results that

there is considerable potential for development of papain enzyme to explore the product

commercialisation at large in Sabah. In summary, it may be stated that spurred by the

phenomenal success of papain enzyme extraction, the industry is actively pursuing

various other applications. Many papain new processes are likely to be commercialised

in the years ahead.

0

20

40

60

80

100

120

1990

1995

2000

2005

Figure 3: Output and projections of papaya in Malaysia during 1990-2005

REFERENCES

Arnon, R. 1970. Papain, in Methods in Enzymology, 19, Academic Press.New York. 220

p.

Blumberg, S., Schechter, I. and Berger, A. 1970. The purification of papain by affinity

chromatography, Eur. J. Biochem 15, 97 p.

Crueger, M. and Crueger, A. 1984. Biotechnology: A textbook of industrialmicrobiology.

Science Tech., Madison, 396 p.

Ensminger, D. 1988. Ultrasonics: Fundamentals, Technology,Applications, 2nd Ed.,

Marcel Dekker, New York.

Harlad, W. T. 1997. Papaya as Medicine: A safe and cheap form of food therapy,Pelanduk Publications (M) Sdn.Bhd. Kau Lampur.


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Kirschenbau, D. 1971. The enhancement of the enzymatic activity of papain by reaction

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Mukherjee, K.D. and Kiewitt I. 1996. Specificity of Carica papaya latex as biocatalyst in,

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Pietsch, W. 1996. Successfully use agglomeration for size enlargement. Chem. Eng.

Progress, 92:29-45.

Ram, M. and Singh, B.N. 1996. Improving productivity, quality and product potential of

papaya. In Vijaysegaran, S. (Eds.) Proc. of the international conference on tropicafruits

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