Abstract
Several unripe fruits of Carica papaya drop in various orchards and farms hence contributing to environmental burden and wastage. The nutritive and ethno-medicinal potentials (proximate, mineral, vitamins and phytochemicals) of the unripe seed in livestock production were evaluated using AOAC methods. Data were analysed using means and standard deviation. The proximate evaluation of the unripe seeds of Carica papaya showed that it contained crude protein (8.90±0.28%), crude fibre (29.00±1.41%), crude fat (29.50±2.12%), ash (8.65±0.64%),
carbohydrates (23.95±4.45%) and moisture (5.45±0.21%). The mineral contents were calcium (0.39±0.00%), magnesium (0.13±0.00%), potassium (0.11±0.00%), sodium (0.04±0.00%), phosphorus (5.71±1.41 mg/kg), manganese (26.91±1.41 mg/kg) and iron (105±1.41 mg/kg). The vitamins contents obtained were 2162.50±1.41 IU/kg, 1.27±0.00 mg/100g, 0.64±0.00 mg/100g, 3.57±1.41 mg/100g, 1.94±1.41 mg/100g, 0.86±0.00 mg/100g and 8.99±1.41 mg/100g for Vitamins A, B1, B2, B3, B6, B12 and C respectively. The values of the phytochemicals were 7.42±1.41 mg/100g, 5.94±1.41 mg/100g, 0.31±0.00 mg/100g and 84.12±1.41 mg/100g for saponin, alkaloid, hydrogen cyanide and tannin respectively. Thus, the unripe seeds of Carica papaya could be a reliable source for minerals, vitamins, fibre, fat and carbohydrate with broader activity and higher potentials for therapeutic. Harnessing the results of this study may enhance the use of these samples in diets and phytobiotics to ultimately reduce their environmental burden.
CHAPTER ONE
• INTRODUCTION
• BACKGROUND OF STUDY
The Carica papaya plant belongs to the Caricaceae family, it is commonly called papaya. This plant is also known in different part if the world as papaw, paw-paw, kapaya, lapaya, tapaya, papayao, papaya, papaia, papita, lechosa, fruta bomba, mamon, mamona, mamao and tree melon. It is native to the Caribbean Coast of Central America (Milind and Gurditta, 2011). Carica papaya is cultivated in tropical and subtropical regions of the world (Lavanya et al., 2018). Papaya is cultivated in the tropical and neo-tropical regions of the world between 32º North and South. Amongst the 31 species of the botanical family Caricaceae and the genera Carica, papaya specie is the most economically important and widely cultivated specie. Carica papaya is the most economically important and widely cultivated species amongst the 31 species of its botanical family (Nur, 2010).
During the early years of growth, papaya develops a single stem; with time in highly fertile soil, thereby promoting favourable growth condition, also heavy lateral branch develops. The leaves of a mature papaya plant are palmate with deep lobes, supported by smooth, hollow petioles (Nur, 2010). The stems, fruits and leaves contain copious amount of latex. It is crowded by terminal cluster of large and long stalked leaves, which rapidly grows and up to 20m of height (Banerjee, 1986).
Papaya species exist in three sex types: male, hermaphrodite and female. In commercial production, the hermaphrodite specie is most desired. A single gene having at least three alleles controls the sex of the plant; a dominant allele for male plants, another dominant allele for hermaphrodite plants and a recessive allele for female plants.
Ripe papaya fruits have slight resemblance with melons; they are rich in retinol and ascorbic acid. They are rich sources of β-carotene, vitamin C, vitamin A and vitamin E, which are highly potent antioxidants; they possess minerals such as calcium, zinc, magnesium, manganese, potassium and iron; they contain also the phytochemicals and the B vitamins (Aravind et al., 2013). Papaya is a good source of fiber. All the nutrients of papaya as a whole improve cardiovascular system, protect against heart diseases, heart attacks, strokes and prevent colon cancer and its enzymes are used in the treatment of arthritis (Milind and Gurditta, 2011). Papaya contains latex present in its unripe fruits as well the leaves and stems. Carica papaya contains biologically active compounds and enzymes (Tigist et al., 2016).
Enzyme benefits the human health. Enzyme was first named in the late 19th century by Wurtz and Bouchut (1945) who purified partially the product from the sap of papaya; it was recognized as a constituent in the latex of tropical papaya fruit when named. From papaya latex, the endolytic plant cysteine protease enzyme – papain and chymopapain is isolated (Amri and Mamboya, 2012). Latex of the papaya contains a sulfhydryl protease ‒papain and chymopapain (Lavanya et al., 2018). Papain (EC 3.4.22.2) and chymopapain are the most common important enzymes of papaya (Milind and Gurditta, 2011). Papain is of vital importance in biological processes in living organisms, it belongs to the papain superfamily, as it is has inherent proteolytic properties (Tsuge et al., 1999). Papain basically is obtained by making incisions on the epicarp of unripe papaya, collecting and drying the latex which flows out. More active papain is gotten from a greener fruit (Amri and Mmaboya, 2012).
According to Menard et al., (1990), papain preferentially cleaves peptide bonds involving basic amino acids, particularly arginine, lysine and residues following phenylalanine. In the 1990s were the precursors and inhibitors of papain studied (Vernet, 1991). The functionality of papain is due to its unique structure which helps in the comprehension of the mechanism of the proteolytic enzyme and its usefulness for a variety of purposes (Amri and Mmaboya, 2012). Papain and chymopapain is soluble in water, their solution has good temperature stability; the solution stability has high dependence on the pH. Papain solutions are unstable under acidic condition such as pH 2.8 and will result in significant loss of activity; they are however stable at a between a pH of 5.5 - 5.9 (Nur, 2010).
Papain is utilized in food processing especially in the tenderization of meat. The protease has the ability to dissolve dead tissue without damaging any living cell; it is used as commercial meat tenderizers. Papaya fruit and papain are used in wound care. Papain is a digestive enzyme of papaya that effectively treats the causes of trauma, allergies and sports injuries (Tigistet al., 2016). Papain has high proteolytic activity and it is applied extensively in the fields of food and medicine. According to Chaudhari (1996), some studies indicate the enzyme helps in the prevention of diabetic-dependent-heart-disease. Papaya leaves traditionally are wrapped around meat and act a tenderizer. Due to the efficiency and less destructive nature of the proteolytic enzymes – papain and chymopapain, when compared to other proteases is the commonly used enzyme in cell isolation procedures on certain tissues (Nur, 2010). Papain is a constituent of contact lens solution. Papain is used in clarifying beer, synthesis of chewing gum and in pharmaceutical industry as digestive medicine. In a study by Huet et al., (2006), traditional medicine through the administration of papain and chymopapain extract from papaya leaves is shown to be effective against the nematodes as papain causes damage to the cuticles of the nematodes. The combination of papain with other proteolytic enzyme is useful in enzyme- therapy for cancer treatment.
• STATEMENT OF RESEARCH PROBLEM
Papain enzyme generally is a sulfylhudryl protease obtained from Carica papaya. Several research carried out on the enzymes of the papaya plant focus on its extraction and application. The enzyme functions in the tenderization of flesh and also enhances the healing of wound. For those who experience poor wound healing will not have to depend on other materials that are expensive, sophisticated which might not be readily available. There are a number of papaya trees on the campus of Mountain Top University (MTU). The present study focuses on the enzymes in the leaf of the tree on the campus.
• AIM AND OBJECTIVES OF STUDY
The aim of this study is to extract, purify and characterize papain enzyme from the leaves of Carica papaya.
The specific objectives of this study are:
• To extract papain from Papaya leaves in MTU.
• To purify the extracted papain.
• To determine the papain concentration at each phase of purification.
• To determine the enzyme activity at each purification phase.
• To characterize the purified papain.
• SCOPE OF STUDY
In order to achieve the objectives, the following procedure were taken:
• Grinding the leaves in a mortar with pestle followed by homogenization in a warring blender.
• Precipitation of the protein with 70% ammonium sulphate solution.
• Dialysis of the homogenate against distilled water.
• Passing the dialysate through DEAE-Cellulose column chromatography.
• Gel filtration of the fractions from DEAE-Cellulose column.
• Determination of the molecular weight, the effect of pH, temperature, substrate concentration and Mg2+ on the enzyme.
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