Fish Physiology and Biochemistry

, Volume 36, Issue 3, pp 491–499 | Cite as

Modulation of key enzymes of glycolysis, gluconeogenesis, amino acid catabolism, and TCA cycle of the tropical freshwater fish Labeo rohita fed gelatinized and non-gelatinized starch diet

  • Vikas Kumar
  • N. P. Sahu
  • A. K. Pal
  • Shivendra Kumar
  • Amit Kumar Sinha
  • Jayant Ranjan
  • Kartik Baruah


A 60-day experiment was conducted to study the effect of dietary gelatinized (G) and non-gelatinized (NG) starch on the key metabolic enzymes of glycolysis (hexokinase, glucokinase, pyruvate kinase, and lactate dehydrogenase), gluconeogenesis (glucose-6 phosphatase and fructose-1,6 bisphosphatase), protein metabolism (aspartate amino transferase and alanine amino transferase), and TCA cycle (malate dehydrogenase) in Labeo rohita juveniles. In the analysis, 234 juveniles (2.53 ± 0.04 g) were randomly distributed into six treatment groups each with three replicates. Six semi-purified diets containing NG and G cornstarch, each at six levels of inclusion (0, 20, 40, 60, 80, and 100) were prepared viz., T1 (100% NG, 0% G starch), T2 (80% NG, 20% G starch), T3 (60% NG, 40% G starch), T4 (40% NG, 60% G starch), T5 (20% NG, 80% G starch), and T6 (0% NG, 100% G starch). Dietary G:NG starch ratio had a significant (P < 0.05) effect on the glycolytic enzymes, the highest activities were observed in the T6 group and lowest in the T1 group. On the contrary, the gluconeogenic enzymes, the glucose-6-phosphatase and fructose-1,6 bisphosphatase activities in the organs, liver and kidney were recorded highest in the T1 group and lowest in the T6 group. The liver aspartate amino transferase activity showed an increasing trend with the decrease in the dietary G level. However, the muscle aspartate amino transferase activity was not significantly (P > 0.05) influenced by the type of dietary starch. The alanine amino transferase activity in both liver and muscle showed an increasing trend with the decrease in the dietary G level. The liver and muscle malate dehydrogenase activities were lowest in the T6 group and highest in the T1 group. Results suggest that NG (100%) starch diet significantly induced more the enzyme activities of amino acid metabolism, gluconeogenesis, and TCA cycle, whereas partial or total replacement of raw starch by gelatinized starch increased the glycolytic enzyme activity.


Starch Labeo rohita Hexokinase Glucokinase Pyruvate kinase Lactate dehydrogenase Glucose-6 phosphatase Fructose-1,6 bisphosphatase Aspartate amino transferase Alanine amino transferase Malate dehydrogenase 



The authors are grateful to the Director, Central Institute of Fisheries Education, Mumbai, for providing facilities for carrying out the work. The first author is grateful to the Central Institute of Fisheries Education, Mumbai, for awarding the institutional fellowship.


  1. Alexis MN, Papaparaskeva-Papoutsoglou E (1986) Aminotransferase activity in the liver and white muscle of Mugil capito fed diets containing different levels of protein and carbohydrate. Comp Biochem Physiol 83B:245–249Google Scholar
  2. Bergot F (1993) Digestibility of native starch of various botanical origins by rainbow trout (Oncorhynchus mykiss). In: Kaushik SJ, Luquet P (eds) Fish nutrition in practice. Proceedings of the IV international symposium on nutrition and feeding, Les Colloques, INRA, no. 61, pp 857–865Google Scholar
  3. Bergot F, Bréque J (1983) Digestibility of starch by rainbow trout: effects of the physical state of starch and of the intake level. Aquaculture 22:81–96. doi: 10.1016/0044-8486(81)90135-6 CrossRefGoogle Scholar
  4. Catacutan MR, Coloso RM (1998) Growth of juvenile Asian sea bass, Lates calcarifer, fed varying carbohydrate and lipid levels. Aquaculture 149:137–144. doi: 10.1016/S0044-8486(96)01432-9 CrossRefGoogle Scholar
  5. Cho CY, Kaushik SJ (1990) Nutritional energetics in fish: energy and protein utilization in rainbow trout (Salmo gairdneri). In: Bourne GH (ed) Aspects of food production, consumption and energy values. World Rev Nutr Diet, Karger, Basel, 61:132–172Google Scholar
  6. Couto A, Enes P, Peres H, Oliva-Teles A (2008) Effect of water temperature and dietary starch on growth and metabolic utilization of diets in gilthead sea bream (Sparus aurata) juveniles. Comp Biochem Physiol A 151:45–50. doi: 10.1016/j.cbpa.2008.05.013 CrossRefGoogle Scholar
  7. Enes P, Panserat S, Kaushik S, Oliva-Teles A (2006) Effect of normal and waxy maize starch on growth, food utilization and hepatic glucose metabolism in European sea bass (Dicentrarchus labrax) juveniles. Comp Biochem Physiol 143A:89–96CrossRefGoogle Scholar
  8. Enes P, Panserat S, Kaushik S, Oliva-Teles A (2008) Hepatic glucokinase and glucose-6-phosphatase responses to dietary glucose and starch in Gilthead Sea bream (Sparus aurata) juveniles reared at two temperatures. Comp Biochem Physiol A 149:80–86. doi: 10.1016/j.cbpa.2007.10.012 CrossRefGoogle Scholar
  9. FAO (2001) FAO yearbook on fishery statistics. Rome, ItalyGoogle Scholar
  10. Fernandez F, Miquel AG, Cordoba M, Varas M, Metón I, Caseras A, Baanante IV (2007) Effects of diets with distinct protein-to-carbohydrate ratios on nutrient digestibility, growth performance, body composition and liver intermediary enzyme activities in gilthead sea bream (Sparus aurata, L.) fingerlings. J Exp Mar Biol Ecol 343:1–10. doi: 10.1016/j.jembe.2006.10.057 CrossRefGoogle Scholar
  11. Fiske CH, Subbarow Y (1925) The colorimetric determination of phosphorus. J Biol Chem 66:375–400Google Scholar
  12. Freeland RA, Harper AL (1959) The study of metabolic pathway by means of adaptation. J Biol Chem 234:1350–1354Google Scholar
  13. Furuichi M, Yone Y (1982) Change in activities of hepatic enzymes related to carbohydrate metabolism of fishes in glucose and insulin-glucose tolerance tests. Bull Jpn Soc Sci Fish 48:463–466CrossRefGoogle Scholar
  14. Fynn-Aikins K, Hung SSO, Hughes SG (1993) Effects of feeding a high level of d-glucose on liver function in juvenile white sturgeon (Acipenser transmontanus). Fish Physiol Biochem 12:317–325. doi: 10.1007/BF00004416 CrossRefGoogle Scholar
  15. Gaye-Siessegger J, Focken U, Becker K (2006) Effect of dietary protein/carbohydrate ratio on activities of hepatic enzymes involved in the amino acid metabolism of Nile tilapia, Oreochromis niloticus (L.). Fish Physiol Biochem 32:275–282. doi: 10.1007/s10695-006-9000-1 CrossRefGoogle Scholar
  16. Guraya HS, Toledo RT (1993) Determining gelatinized starch in a dry starchy product. J Food Sci 58:888. doi: 10.1111/j.1365-2621.1993.tb09384.x CrossRefGoogle Scholar
  17. Hilton JW, Slinger SJ (1983) Effect of wheat bran replacement of wheat midlings in extrusion processed (floating) diets on the growth of juvenile rainbow trout (Salmo gairdneri). Aquaculture 5:201–210. doi: 10.1016/0044-8486(83)90091-1 CrossRefGoogle Scholar
  18. Hung SSO, Fynn-Aikins FK, Lutes PB, Xu R (1989) Ability of juvenile white sturgeon (Acipenser transmontanus) to utilize different carbohydrate sources. J Nutr 119:727–733CrossRefGoogle Scholar
  19. Hutchins CG, Rawles SD, Gatlin DMIII (1998) Effects of dietary carbohydrate kind and level on growth, body composition and glycemic response of juvenile sunshine bass (Morone chrysops female × M. saxatilis male). Aquaculture 161:187–199. doi: 10.1016/S0044-8486(97)00269-X CrossRefGoogle Scholar
  20. Jeong KS, Takeuchi T, Okamoto N, Watanabe T (1992a) The effects of dietary gelatinized ratios at different dietary energy on growth and characteristic of blood in rainbow trout fingerlings. Nippon Siusan Gakkaishi 58:937–944CrossRefGoogle Scholar
  21. Jeong KS, Takeuchi T, Okamoto N, Watanabe T (1992b) The effect of dietary gelatinized rations at different dietary energy levels on growth and characteristics of blood in carp fingerlings. Bull Jpn Soc Sci Fish 58:945–951CrossRefGoogle Scholar
  22. Kayne F (1973) Pyruvate kinase, the enzymes, 3rd edn. Academic Press, New York, p 353Google Scholar
  23. Kumar S, Sahu NP, Pal AK, Choudhury D, Mukherjee SC (2006) Studies on digestibility and digestive enzyme activities in Labeo rohita (Hamilton) juveniles: effect of microbial α–amylase supplementation in non-gelatinized or gelatinized corn based diet at two protein level. Fish Physiol Biochem 32:209–220. doi: 10.1007/s10695-006-9002-z CrossRefGoogle Scholar
  24. Kumar V, Sahu NP, Pal AK, Kumar S (2007) Immunomodulation of Labeo rohita juveniles due to dietary gelatinized and non-gelatinized starch. Fish Shellfish Immunol 23:341–353. doi: 10.1016/j.fsi.2006.11.008 CrossRefGoogle Scholar
  25. Kumar V, Sahu NP, Pal AK, Kumar S, Gupta SK (2008a) Gelatinized to non-gelatinized starch ratio in the diet of Labeo rohita: effect on digestive and metabolic response and on growth. J Anim Physiol Anim Nutr (Berl) 92:492–501. doi: 10.1111/j.1439-0396.2007.00739.x CrossRefGoogle Scholar
  26. Kumar S, Sahu NP, Pal AK, Sagar V, Sinha AK, Baruah K (2008b) Modulation of key metabolic enzyme of Labeo rohita (Hamilton) juvenile: effect of dietary starch type, protein level and exogenous a-amylase in the diet. Fish Physiol Biochem. doi: 10.1007/s10695-008-9213-6. (in press)CrossRefPubMedGoogle Scholar
  27. Lee DJ, Putnam GB (1973) The response of rainbow trout to varying protein/energy ratios in a test diet. J Nutr 103:916–922CrossRefGoogle Scholar
  28. Marjorie AS (1964) In: Colowick SP, Kaplan NO (eds) Methods in enzymology, vol II. Academic Press Inc, New York, p 541Google Scholar
  29. Mohapatra M, Sahu NP, Chaudhari A (2002) Utilization of gelatinized carbohydrate in diets in Labeo rohita fry. Aquacult Nutr 8:1–8. doi: 10.1046/j.1365-2095.2002.00176.x CrossRefGoogle Scholar
  30. Ochoa S (1955) Malic dehydrogenase and ‘malic’ enzyme. In: Coloric SP, Kaplan N (eds) Methods of enzymology I. Academic Press, New York, pp 735–745CrossRefGoogle Scholar
  31. Peres MH, Oliva-Teles A (2002) Utilization of raw and gelatinized starch by European sea bass (Dicentrarchus labrax) juveniles. Aquaculture 205:287–299. doi: 10.1016/S0044-8486(01)00682-2 CrossRefGoogle Scholar
  32. Rosas C, Cuzon G, Gaxiola G, LePriol Y, Pascual C, Rossignyiol J, Contreras F, Sanchez A, Van Wormhoudt A (2001) Metabolism and growth of juveniles of L. vannamei: effect of salinity and dietary carbohydrates level. J Exp Mar Biol Ecol 259:1–22. doi: 10.1016/S0022-0981(01)00222-2 CrossRefGoogle Scholar
  33. Small BC, Soares JH Jr (1999) Effect of dietary carbohydrate on growth, glucose tolerance and liver composition of juveniles striped bass. N Am J Aquaculture 61:286–292. doi: 10.1577/1548-8454(1999)061<0286:EODCOG>2.0.CO;2 CrossRefGoogle Scholar
  34. Suárez MD, Sanz A, Bazoco J, García-Gallego M (2002) Metabolic effects of changes in the dietary protein: carbohydrate ratio in eel (Angilla anguilla) and trout (Oncorhynchus mykiss). Aquacult Int 10:143–156. doi: 10.1023/A:1021371104839 CrossRefGoogle Scholar
  35. Tranulis MA, Dregni O, Christophersen B, Krogdahl A, Borrebaek B (1996) A glucokinase-like enzyme in the liver of Atlantic salmon (Salmo salar). Comp Biochem Physiol 114B:35–39CrossRefGoogle Scholar
  36. Tung PH, Shiau SY (1993) Carbohydrate utilization versus body size in tilapia Oreochromis niloticus × O. aureus. Comp Biochem Physiol A 104:585–588. doi: 10.1016/0300-9629(93)90468-J CrossRefGoogle Scholar
  37. Verma AK, Pal AK, Manush SM, Das T, Dalvi RS, Chandrachoodan PP, Ravi PM, Apte SK (2007) Persistent sub-lethal chlorine exposure elicits the temperature induced stress responses in Cyprinus carpio early fingerlings. Pestic Biochem Physiol 87:229–237. doi: 10.1016/j.pestbp.2006.08.001 CrossRefGoogle Scholar
  38. Walton MJ (1986) Metabolic effects of feeding a high protein/low carbohydrate diet as compared to a low protein/high carbohydrate diet to rainbow trout Salmo gairdneri. Fish Physiol Biochem 1:7–15. doi: 10.1007/BF02309589 CrossRefGoogle Scholar
  39. Walton MJ, Cowey CB (1982) Aspects of intermediary metabolism in salmonid fish. Comp Biochem Physiol 73B:59–79Google Scholar
  40. Wilson RP (1994) Utilization of dietary carbohydrate by fish. Aquaculture 124:67–80. doi: 10.1016/0044-8486(94)90363-8 CrossRefGoogle Scholar
  41. Wilson RP, Poe WE (1987) Apparent inability of channel catfish to utilize dietary mono- and disaccharides as energy sources. J Nutr 117:280–285CrossRefGoogle Scholar
  42. Wootton IDP (1964) Enzymes in blood. In: Churchill J, Churchill A (eds) Microanalysis in medical biochemistry, 4th edn. Churchill, London, UK, pp 101–107Google Scholar
  43. Wroblewski L, Ladue JS (1955) LDH activity in blood. Proc Soc Exp Biol Med 90:210–213CrossRefGoogle Scholar
  44. Yengkokpam S, Sahu NP, Pal AK, Mukherjee SC, Debnath D (2006) Gelatinized carbohydrates in the diet of Catla catla Fingerlings: effect of levels and sources on nutrient utilization, body composition and tissue enzyme activities. Asian-australas J Anim Sci 20:89–99CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Vikas Kumar
    • 1
  • N. P. Sahu
    • 2
  • A. K. Pal
    • 2
  • Shivendra Kumar
    • 2
  • Amit Kumar Sinha
    • 3
  • Jayant Ranjan
    • 3
  • Kartik Baruah
    • 3
  1. 1.Department of Aquaculture System and Animal Nutrition in the Tropics and Subtropics (480b)University of HohenheimStuttgartGermany
  2. 2.Division of Fish Nutrition and BiochemistryCentral Institute of Fisheries Education, VersovaMumbaiIndia
  3. 3.Laboratory of Aquaculture and Artemia, Department of Animal ProductionGhent UniversityGhentBelgium

Personalised recommendations