Fruit Pulp and Peel.During Ripening of White - and Pink-Fleshed Guava Fruit Types Were Investigated
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'Kind'K A bdclittonetn rBasfiir - - B.Sc..(Agile.) Honours (Khartoum) A Thesis Submitted in partial fulfilment of the requirement Tor (lie degree of Master of Science (Agric.) Supervisor: Dr. Abu-Bakr Ali Abu-Goukh MSPARTMICNT OF HOUTICUI/I'UKIi FACULTY OF AGRICULTURE UNIVERSITY OF KHARTOUM MAY, 1999 3 1/28 Please be aware that all of the Missing Pages in this document were originally blank pages (In the name of Allah, the Beneficent, the Merciful) My Lord: "Increase me in 1(iiozi/(ed(je" (20:114, Ta Ha, Holly Quran) Acknowledgment My special praise and unlimited thanks avtto ALLAH; who supplied helped and gave me health and patience to complete this study. I wish to express my deep sense ol' gratitude and sincere thanks to my supervisor Dr. Abu-Bakr AH Abu-Golikh for his encouragement, excellent suggestions, continued interest and guidance during my study. My greal thanks to Dr. Farouk Hassan El Tahir, Dr. Mustafa ElbalJa hn~ continuous interest, help and encouragement. Thanks and also due to the members of Horticulture and Bioclieuisir_y_De4iartmeut, I am grateful to my family members for their help and encoruagemenl. (ii) ABSTRACT Compositional changes in fruit pulp and peel.during ripening of white - and pink-fleshed guava fruit types were investigated. Fruit tissue firmness decreased progressively, in a simMar manner, in both guava types. The white and pink guavas exhibited a typical climacteric pattern of respiration, with.climacteric peak al 7.6 kilogram-force (kg. f) flesh firmness in both types. Total soluble solids (TSS) and total sugars increased in pulp and peel of both guava types with decrease in flesh firmness.. More increase in total sugars was observed after the climacteric peak. Reducing sugars and titratable acidity increased up to the climacteric peak and decreased afterwards. Total protein increased up to the -fttthripe stage- antHftmr-decreiised. Asco^ compounds decreased continuously during ripening of the two types. The peel showed higher values of ascorbic acid and phenolic compounds compared to the pulp. The white-fleshed guavas had higher levels of TSS, total sugars, reducing sugars, titralable acidity and ascorbic acid content compared to the pink-fleshed fruits. Changes in the activities of the cell wall degrading enzymes, pectinesterase (PE), polygalacluronase (PG) and cellulasc were also studied to find out the reason for tissue softening in g.uava fruit during handling, transportation and storage. (ii) Pectinesterase activity, in both guava types, increased gradually till the climacteric peak and then decreased. Polygalacturonase and cellulase activities progressively increased during ripening and high correlation was observed between the increase in enzyme activity and the loss in fruit flesh firmness. It was concluded'.that PE, PG and cellulase enzymes play an important role in controlling tissue softening mechanism during fruit ripening in guavas. (iii) CONTENTS Page Abstract - .- ii Arabic Abstract -. iv List of Figures ix Chapter 1 : Introduction I Chapter 2: Literature Review - 3 J2..I. Origin, Production and Distribution -• 3 2 .2. Fruit Characteristic 3 2.3. Fruit Ripening and Senescence ----- 4 2 .4. Respiration Rale 5 % .5. Compositional Changes During Fruit Ripening 7 J2.5.1. Total Soluble Solids 7 -.2.5.2. Total and Reducing Sugars 7 2. 5.3. Acidity and pH 8 2 • 5.4. Amino Acids and Proteins 9 2. «5.5. Phenolic Compounds 10 2. • 5.6. Vitamins and Minerals 11 2 .6. Fruit Softening - 13 £•6.1. Role of Pcctinestcrase 15 £.6.2. Role of Polygalacturonase 16 £.6.3. RoleofCellulase 18 (vi) Page Chapter 3 : Materials and Methods 20 3.1. Fruit Source and Ripening Procedure 20 3.2. Respiration and Firmness Determination .- 20 3.3. Total Soluble Solids Determination 20 3.4. Compositional Changes 21 3.4.1. Tissue Extraction 21 3.4.2. Titralable Acidity Determination 21 3.4.3. Total Sugars Determination 21 3.4.4. Reducing5Tugars Determination 21 3.4.5. Total proteins Determination - 22 3.4.6. Phenolic Compounds Determination 'l'l 3.4.7. Ascorbic/I cid Determination •-•- 22 3.5. Enzymatic Activity Changes - 22 3.5.1. Enzyme Extraction • 22 3.5.2. Pectineslerase Activity Determination 23 3.5.3. Polygalacturonase Activity Determination 24 3.5.4. Cellulase Activity Determination 24 Chapter 4: Results 25 .4.1. Fruit Firmness and Respiration Rate. - 25 4.2. Compositional Changes During Ripening in Guava Fruit 25 4.2.1. Total Soluble Solids 25 (vii) Page 4.2.2. Total Sugars - 26 4.2.3. Reducing Sugars - 26 4.2.4. Titratable Acidity 27 4.2.5. Total Proteins 27 4.2.6. Phenolic Compounds 27 4.2.7. Ascorbic Acid 28 4.3. Enzymatic Activily(^hanges During Ripening of Guava fruit 28 4.3.1. Pectinesterase * 29 4.3.2. Polygalacluronasc 29 4.3.3. Cellulase -' 29 Chapter 5 : Discussion 43 Conclusion - 55 References 56 Appendixes - - 74 (viii) List u£ Figures Page Figure : 1. Changes of fruit flesh firmness during ripening of while-and pink-fleshed guava fruits. 31 2. Relationship between CO2 production and fruit flesh firmness during ripening of white-and pink-fleshed guava fruits 32 3. Changes of total soluble solids (TSS) during ripening of white-and pink-fleshed guava fruits 33 4. Total sugars changes during ripening of vvhitc-and pink-fleshed guava fruits. - 34 5. Reducing sugars changes during ripening of white-and pink-fleshed guava fruits ---• .35 6. Changes in titratablc acidity during ripening of while-and pink-fleshed guava fruits 36 "?. ^TaTT0^nnntirhfjfinTn"n"cioTiteTit daring ripening of "white-and pink-fleshed guava fruits 37 - 8. Changes in phenolic compounds during ripening oi' whilc-and pink-fleshed guava fruit 38 9. Changes in ascorbic acid content during ripening of white-and pink-fleshed guava fruits - 39 10. Changes in PE activity d^crign ripening of white-and pink-fleshed guava fruits 40 11. Changes in PG activity during ripening of white-and pink-fleshed guava fruits 41 12. Changes in cellulase activity during ripening of'white-and ' pink-fleshed guava fruits 42 (ix) CHAPTER 1 CHAPTER 1 INTRODUCTION Guava {psidium guajava L.) is native of tropical countries. It can grow satisfactory even in adverse environmental conditions. The fruit has a characteristic odour and is eaten fresh, processed or cooked (Saiunkhe and Desai, 1984). Ripe guava fruit is an excellent source of ascorbic acid (vitamin C) and dietary fiber. It has about five times more vitamin C than orange (guava, 242 mg. per lOOg. j orange, 50mg. per lOOg.) (Watt and Merrill, 1975). Guava is also a good source of vitamin A, phosphorus, calcium and iron as well as tln'amin and niacin (Wilson, -1-986)— Guava in Sudan is still a minor fruit crop, but it is commercially' grown in every region in the Country. Sudan has great potentiality to produce good Quality guava fruits. It has good marketability in the middle east and European countries. The fruit is delicate and many culttvars cannot withstand long-distance transportation and may reach the market in a mushy, over-ripe state. It can store for only few days at ambient conditions. Ripening of the fruit is characterized-by different compositional changes and by softening of the flesh. Development of proper handling techniques and control of the ripening proc'ess are of crucial importance for development of a sound guava industry in the Sudan. This study was carried out with the following objectives: 1. To provide base-line information regarding the biochemical nature of ripening and softening mechanisms in the guava fruit, to assist in development of sound programmes for controlling fruit ripening and loss of firmness during handling, transportation and storage of guava fruits. 2 - To study compositional changes in pulp and peel of white - and pink-fleshed guavas during fruit ripening. 3. To investigate changes in the activity of the cell wall degrading enzymesjpcclincstcrasc (PE), poiygalacturonasc (PG), and cellulase during guava fruit ripening. CHAPTER 2 LITERATURE REVIEW 2..1- Origin, Production and Distribution: Guava (Psidium guajava L.) is an important cultivated species of the Myrtle family (Salunk he and Desai, 1984). It is native to the tropical regions of America. It was spread first into the Philippines and India, and then quickly extended to most of the tropical and subtropical regions of the world, _where it has become naturalized (Yadav, 1998). The crop tolerates a wide range of climates and is fairly salt and drought resistant. There could be 2 to 3 harvest seasons in one.year. It has been reported that fivc-ycar-old guava trees can produce from 130 to 225 kilograms of fruit per tree (28 to 48 tons per Guava is a minor fruit crop. The major producing countries arc :• India. (200,000), Mexico (127,000), Egypt (34,000) and Venezuela (17,000) metric tons (Anon, 1978). , In Sudan, guava is a popular fruit crop. It is grown in every slate. Marketability is limited to local markets due to the delicate nature of the fruit and poor transportation and storage facilities. 2 »2. Fruit characteristics: Guava fruit has a rough yellow skin and is round, ovoid, or pyriform shape. The flesh colour varies from white, deep pink to salmon red, and the flavour has been described as sweet, mushy, and highly aromatic. The fruit is eaten fresh, processed or cooked. It is used for the preparation of juices, jams, jellies, pastes and other similar products (Salunkhe and Desai, 1984). Guava fruit is very rich in ascorbic acid (Wilson, 1980). It is a fair source of beta-carotene (280 IU), calcium (23mg), phosphorous (42mg), riboflavin (0.05'mg), thiamine (0.05mg) and niacin (1.2mg per lOOg) (Watt and Merrill, 1975). Reportedly, consumption • of'guava fruit reduces serum cholesterol levels, triglycerides and hypertension, and increases the level of high density lipoprotein.