Indian Journal of Biochemistry 
& Biophysics

 

Total visitors: 3,720  since 19-10-06

 

VOLUME 43

NUMBER 5

OCTOBER 2006

 

CONTENTS

Minireview

 

Hypolactasia as a molecular basis of lactose intolerance

267

Kamaljit Kaur, Safrun Mahmood and Akhtar Mahmood*

 

 

Biochemistry of homocysteine in health and diseases

275

S Ramakrishnan*, K N Sulochana, S Lakshmi, R Selvi and N Angayarkanni

 

 

Papers

 

Characterization of carotenoid hydroxylase gene promoter in
Haematococcus pluvialis

284

      C X Meng, W Wei, Z-L Su and S Qin*

 

 

 

Purification and characterization of trehalose-6-phosphate synthase from Saccharomycopsis fibuligera A11

289

      Likun Liang, Zhenming Chi*, Lingmei Gao and Liyan Ma

 

 

 

Isolation and identification of α-amylase producing Bacillus sp. from dhal industry waste

295

      S Thippeswamy, K Girigowda and V H Mulimani*

 

 

 

Hepatoprotective effect of aqueous extract of Phyllanthus niruri on nimesulide-induced oxidative stress in vivo

299

      Mary Chatterjee and Parames C Sil*

 

 

 

Protective effects of silymarin, a milk thistle (Silybium marianum) derivative on ethanol-induced oxidative stress in liver

306

      Subir Kumar Das* and D M Vasudevan

 

 

 

Photoilluminated riboflavin/riboflavin-Cu(II) inactivates trypsin: Cu(II) tilts the balance

312

      E Husain, R A Fatima, I A F Ali and I Naseem*

 

 

 

Notes

 

Identification of α-2u globulin in the rat preputial gland by MALDI-TOF analysis

319

      P Ponmanickam and G Archunan*

 

 

 

Extraction and carrier-facilitated transport of amino acids using synthetic non-cyclic receptors through bulk liquid membrane

323

Pratibha Joshi, Nidhi Joshi and Uma Sharma*

 

 

 

Instructions to Authors

327

 

 

——————

*Author for correspondence

 

 

 

 

Minireviews

 

 

Indian Journal of Biochemistry & Biophysics

Vol. 43, October 2006, pp. 267-274

 

Hypolactasia as a molecular basis of lactose intolerance

 

Kamaljit Kaur, Safrun Mahmood§ and Akhtar Mahmood*

Department of Biochemistry, Panjab University, Chandigarh 160 014

§Department of Experimental Medicine and Biotechnology, Postgraduate Institute of Medical Education and Research,
Chandigarh 160 012, India

Received 23 May 2006; revised 21 July 2006

Lactase-phlorizin hydrolase (LPH), a membrane-bound glycoprotein present in the luminal surface of enterocytes in the intestine is responsible for lactose intolerance, a phenomenon prevalent in humans worldwide. In the rodent intestine, the post-natal development of the LPH follows a specific pattern, such that the enzyme levels are high in the peri-natal period, but declines considerably upon maturation. The observed maturational decline in the LPH activity is very similar to adult-type hypolactasia observed in humans. Majority of the studies have been carried out using animal models or cell lines and a number of hypotheses have been put forward to explain the maturational decline of lactase activity such as: (a) decreased amount of lactase protein, (b) defect in post-translational modification of precursor lactase to the mature enzyme, and (c) synthesis of an inactive, high molecular weight lactase with altered glycosylation, however, the precise underlying mechanism of adult-type hypolactasia remains undefined. The present review describes the recent developments in understanding the regulation of lactase expression and the possible mechanism of adult-type hypolactasia, as a cause of lactose intolerance.

Keywords: Lactase-phlorizin hydrolase, Lactase activity, Adult-type hypolactasia, Lactase glycosylation and regulation, Lactose intolerance, Lactose gene expression

*Email: akhtarmah@yahoo.com

 

 

Indian Journal of Biochemistry & Biophysics

Vol. 43, October 2006, pp. 275-283

 

Biochemistry of homocysteine in health and diseases

 

S Ramakrishnan*, K N Sulochana, S Lakshmi, R Selvi and N Angayarkanni

Biochemistry Research Department, Vision Research Foundation, Sankara Nethralaya, Chennai 600 006, India

Received 13 January 2006; revised 25 August 2006

The amino acid homocysteine (Hcy), formed from methionine has profound importance in health and diseases. In normal circumstances, it is converted to cysteine and partly remethylated to methionine with the help of vit B12 and folate. However, when normal metabolism is disturbed, due to deficiency of cystathionine-b-synthase, which requires vit B6 for activation, Hcy is accumulated in the blood with an increase of methionine, resulting into mental retardation (homocystinuria type I). A decrease of cysteine may cause eye diseases, due to decrease in the synthesis of glutathione (antioxidant). In homocystinurias type II, III and IV, there is accumulation of Hcy, but a decrease of methionine, thus, there is no mental retardation. Homocysteinemia is found in Marfan syndrome, some cases of type I diabetes and is also linked to smoking and has genetic basis too. In hyperhomocysteinemias (HHcys), clinical manifestations are mental retardation and seizures (type I only), ectopia lentis, secondary glaucoma, optic atrophy, retinal detachment, skeletal abnormalities, osteoporosis, vascular changes, neurological dysfunction and psychiatric symptoms. Thrombotic and cardiovascular diseases may also be encountered. The harmful effects of homocysteinemias are due to (i) production of oxidants (reactive oxygen species) generated during oxidation of Hcy to homocystine and disulphides in the blood. These could oxidize membrane lipids and proteins, (ii) Hcy can react with proteins with their thiols and form disulphides (thiolation), (iii) it can also be converted to highly reactive thiolactone which could react with the proteins forming -NH-CO- adducts, thus affecting the body proteins and enzymes. Homocystinuria type I is very rare (1 in 12 lakhs only) and is treated with supplementation of vit B6 and cystine. Others are more common and are treated with folate, vit B12 and in selected cases as in methionine synthase deficiency, methionine, avoiding excess. In this review, the role of elevated Hcy levels in cardiovascular, ocular, neurologial and other diseases and the possible therapeutic measures, in addition to the molecular mechanisms involved in deleterious manifestations of homocysteinemia, have been discussed.

Keywords: Homocysteine metabolism, Homocystinurias, Homocysteinemias, Clinical manifestations, Thrombotic disorders, Cardiovascular diseases, Diabetes, Ocular diseases, Smokers, Oxidant stress, Protein thiolation, Protein homocysteinylation, Cystine, Vitamins B6, B12, folate.

*E mail: drrk@snmail.org

 

 

Papers

 

Indian Journal of Biochemistry and Biophysics

Vol. 43, October 2006, pp. 284-288

 

Characterization of carotenoid hydroxylase gene promoter in
Haematococcus pluvialis

C X Meng1,2#, W Wei1,2, Z-L Su1 and S Qin1*

1Institute of Oceanology, Chinese Academy of Sciences, Qingdao, 266071, China

2Graduate school, Chinese Academy of sciences, Beijing, 100039, China

Received 16 Januaury 2006; revised 6 July 2006

Astaxanthin, a high-value ketocarotenoid is mainly used in fish aquaculture. It also has potential in human health due to its higher antioxidant capacity than β-carotene and vitamin E. The unicellular green alga Haematococcus pluvialis is known to accumulate astaxanthin in response to environmental stresses, such as high light intensity and salt stress. Carotenoid hydroxylase plays a key role in astaxanthin biosynthesis in H. pluvialis. In this paper, we report the characterization of a promoter-like region (–378 to –22 bp) of carotenoid hydroxylase gene by cloning, sequence analysis and functional verification of its 919 bp 5-flanking region in H. pluvialis. The 5-flanking region was characterized using micro-particle bombardment method and transient expression of LacZ reporter gene. Results of sequence analysis showed that the 5-flanking region might have putative cis-acting elements, such as ABA (abscisic acid)-responsive element (ABRE), C-repeat/dehydration responsive element (C-repeat/DRE), ethylene-responsive element (ERE), heat-shock element (HSE), wound-responsive element (WUN-motif), gibberellin-responsive element (P-box), MYB-binding site (MBS) etc., except for typical TATA and CCAAT boxes. Results of 5 deletions construct and β-galactosidase assays revealed that a highest promoter-like region might exist from –378 to –22 bp and some negative regulatory elements might lie in the region from –919 to –378 bp. Results of site-directed mutagenesis of a putative C-repeat/DRE and an ABRE-like motif in the promoter-like region (–378 to –22 bp) indicated that the putative C-repeat/DRE and ABRE-like motif might be important for expression of carotenoid hydroxylase gene.

Keywords: Haematococcus pluvialis, Astaxanthin, Carotenoid hydroxylase, cis-Acting element, Promoter

*E-mail: zq77232000@yahoo.com

 

 

Indian Journal of Biochemistry & Biophysics

Vol. 43, October 2006, pp 289-294

 

Purification and characterization of trehalose-6-phosphate synthase from Saccharomycopsis fibuligera A11

 

Likun Liang, Zhenming Chi*, Lingmei Gao and Liyan Ma

UNESCO Chinese Center of Marine Biotechnology, Ocean University of China, Yushan Road, No.5, Qingdao, China

Received 13 March 2006; revised 2 September 2006

Mutant A11, a mutant of Saccharomycopsis fibuligera Sdu with low acid and neutral trehalase was found to accumulate over 18% (w/w) trehalose from starch in its cells. In this study, trehalose-6-phosphate synthase (Tps1) was purified to homogeneity from this mutant, with a 30-fold increase in the specific enzyme activity, as compared to the concentrated cell-free extract, from initial cells. The molecular mass of the purified enzyme as determined by SDS-PAGE was 66 kDa. The optimum pH and temperature of the purified enzyme were 6.6 and 37°C, respectively. The enzyme was activated by Ca2+, K+ and Mg2+, with K+ showing the highest activation at 35 mM. On the other hand, Mn2+, Cu2+, Fe3+, Hg2+ and Co2+ inhibited the enzyme. The enzyme was also strongly inhibited by protease inhibitors such as iodoacetic acid, EDTA and PMSF.

Keywords: Saccharomycopsis fibuligera, Trehalose-6-phopshate synthase, Enzyme purification, Characterization of Tps1

*E-mail: zhenming@sdu.edu.cn

 

 

 

Indian Journal of Biochemistry & Biophysics

Vol. 43, October 2006, pp. 295-298

 

 

Isolation and identification of α-amylase producing Bacillus sp. from dhal industry waste

S Thippeswamy, K Girigowda and V H Mulimani*

Department of Studies in Biochemistry, Gulbarga University, Gulbarga 585 106, Karnataka, India

Received 14 February 2006; revised 8 August 2006

A bacterial strain was isolated from dhal industry red gram waste and identified as Bacillus. A thermostable extracellular amylase was partially purified from the strain. Optimum temperature and pH for the enzyme were found to be 60°C and 6.5, respectively. The maximum amylase production was achieved with maltose as carbon source. Among the nitrogen sources, peptone and yeast extract produced maximum amylase.

Keywords: Bacillus spp., Thermostable amylase, Dhal industry waste.

*E mail: v_h_mulimani@rediffmail.com

 

 

 


Indian Journal of Biochemistry & Biophysics

Vol. 43, October 2006, pp 299-305

 

Hepatoprotective effect of aqueous extract of Phyllanthus niruri on nimesulide-induced oxidative stress in vivo

 

Mary Chatterjee and Parames C Sil*

Department of Chemistry, Bose Institute, 93/1, Acharya Prafulla Chandra Road Kolkata-700009, India

Received 26 October 2005; revised 5 September 2006

Nimesulide (NIM), an atypical non-steroidal anti-inflammatory drug (NSAID) is also used as analgesic. In the present study, we evaluated its effect on the prooxidant-antioxidant system of liver and the hepatoprotective potential of aqueous extract of the herb Phyllanthus niruri (PN) on NIM-induced oxidative stress in vivo using a murine model, by determining the activities of hepatic anti-oxidant enzymes superoxide dismutase (SOD) and catalase (CAT), levels of reduced glutathione (GSH) and lipid peroxidation (expressed as malonaldialdehyde, MDA). Aqueous extract of PN at a dose of 50 or 100 mg/kg body wt was administered either intraperitoneally or orally for 7 days, before NIM administration at a dose of 8 mg/kg body wt twice daily for 7 days in mice. Animals were sacrificed 24 h after administration of final dose of NIM. In another set of experiments, both aqueous extract of PN (at a dose of 50 or 100 mg/kg body wt) and NIM (8 mg/kg body wt) were administered simultaneously for 7 days. Animals were sacrificed 24 h after administration of final dose of the extract and NIM, liver tissues were collected, and the activities of SOD and CAT and levels of GSH and lipid peroxidation end-product (as MDA), were determined from the livers of all the experimental animals. Appropriate NIM control was maintained for all sets of experiments. NIM administration (8 mg/kg body wt) for 7 days caused significant depletion of the levels of SOD, CAT and reduced GSH, along with the increased levels of lipid peroxidation. Intraperitoneal administration of the extract at a dose of 50 mg/kg body wt for 7 days, prior to NIM treatment, significantly restored most of the NIM-induced changes and the effect was comparable to that obtained by administering 100 mg/kg body wt of the extract orally. Thus, results suggested that intraperitoneal administration of the extract could protect liver from NIM-induced hepatic damage more effectively than oral administration. Antioxidant property of the aqueous extract of PN was also compared with that of a known potent antioxidant, vitamin E. The PN extract at a dose of 100 mg/kg body wt along with NIM was more effective in suppressing the oxidative damage than the PN extract at a dose of 50 mg/kg body wt. Results suggested that beneficial effect of the aqueous extract of PN, probably through its antioxidant property, might control the NIM-induced oxidative stress in the liver.

Keywords: Nimesulide, Oxidative stress, Phyllanthus niruri, Antioxidants, Hepato-protective activity.

*E-mail: parames@bosemain.boseinst.ac.in; parames_95@yahoo.co.in

 

 

 

 

Indian Journal of Biochemistry & Biophysics

Vol. 43, October 2006, pp. 306-311

 

Protective effects of silymarin, a milk thistle (Silybium marianum) derivative on ethanol-induced oxidative stress in liver

Subir Kumar Das* and D M Vasudevan

Department of Biochemistry, Amrita Institute of Medical Sciences, Cochin 682 026, Kerala, India

Received 28 February 2006; revised 17 July 2006

The production of reactive oxygen species (ROS) is considered to be a major factor in oxidative cell injury. The antioxidant activity or the inhibition of the generation of free radicals is important in providing protection against such hepatic damage. Silymarin, derived from the milk thistle plant, Silybium marianum, has been used in traditional medicine as a remedy for diseases of the liver and biliary tract. In the present study, the effect of hepatoprotective drug silymarin on body weight and biochemical parameters, particularly, antioxidant status of ethanol-exposed rats was studied and its efficacy was compared with the potent antioxidant, ascorbic acid as well as capacity of hepatic regeneration during abstention. Ethanol, at a dose of 1.6 g/kg body wt/day for 4 wks affected body weight in 16-18 week-old male albino rats (Wistar strain weighing 200-220 g). Thiobarbituric acid reactive substance (TBARS) level, superoxide dismutase (SOD), and glutathione-s-transferase (GST) activities were significantly increased, whereas GSH content, and catalase, glutathione reductase (GR) and GPx (glutathione peroxidase) activities significantly reduced, on ethanol exposure. These changes were reversed by silybin and ascorbic acid treatment. It was also observed that abstinence from ethanol might help in hepatic regeneration. Silybin showed a significant hepatoprotective activity, but activity was less than that of ascorbic acid. Furthermore, preventive measures were more effective than curative treatment.

Keywords: Silybium marianum, Ascorbic acid, Ethanol, Oxidative stress, Silybin, Silymarin, antioxidant status

 

*E-mail: subirkumardas@aims.amrita.edu

 

 

Indian Journal of Biochemistry & Biophysics

Vol. 43, October 2006, pp. 312-318

 

 

Photoilluminated riboflavin/riboflavin-Cu(II) inactivates

trypsin: Cu(II) tilts the balance

 

E Husain1 R A Fatima1­­, I A F Ali­2 and I Naseem1*

1Department of Biochemistry, Faculty of Life Sciences, Aligarh Muslim University, Aligarh 202 002 (U.P.), India

2Faculty of Pharmacy, Al–Najah National University, Nablus, Palestine

Received 16 January 2006; revised 31 August 2006

Riboflavin (RF) upon irradiation with fluorescent light generates reactive oxygen species like superoxide anion, singlet and triplet oxygen, flavin radicals and substantial amounts of hydrogen peroxide (H2O2). H2O2 can freely penetrate cell membrane and react with a transition metal ion like Cu(II), generating hydroxyl radical via the modified metal-catalyzed Haber-Weiss reaction. Earlier, it was reported that trypsin-chymotrypsin mixture served as an indirect antioxidant and decreased free radical generation. Thus, in the present study, we used photoilluminated RF as a source of ROS to investigate the effect of free radicals on the activity of trypsin. We also compared the damaging effect of photoilluminated RF and RF-Cu(II) system using trypsin as a target molecule. RF caused fragmentation of trypsin and the effect was further enhanced, when Cu(II) was added to the reaction. Results obtained with various ROS scavengers suggested that superoxide radical, singlet and triplet oxygen were predominantly responsible for trypsin damage caused by photoilluminated RF. On the other hand, when Cu(II) was added to the reaction, hydroxyl radical was mainly responsible for trypsin damage. A mechanism of generation of various ROS in the reaction is also proposed. Trypsin did not show any antioxidant effect with RF alone or with RF–Cu(II) combination.

Keywords: Riboflavin, Trypsin, Copper, Reactive oxygen species

*E-mail: imrananaseem@yahoo.com

 

Notes

 

Indian Journal of Biochemistry & Biophysics

Vol. 43, October 2006, pp. 319-322

 

 

Identification of α-2u globulin in the rat preputial gland by MALDI-TOF analysis

P Ponmanickam and G Archunan*

Department of Animal Science, Bharathidasan University
Tiruchirappalli 620 024, TN, India

Received 18 April 2006; revised 13 July 2006

The low molecular mass proteins found in the pheromonal sources such as urine, saliva, glandular secretion etc have been reported as ligand carriers for the processes of chemo-communication in mammals. The preputial gland plays an important role in the production of olfactory signals for pheromonal communication. Thus, in the present study, α-2u globulin having molecular mass of 18 kDa has been identified in the preputial gland of Norway rat (Rattus norvegicus) by in-gel trypsin digestion and analyzing the resulting peptides by MALDI-TOF. Since preputial gland is one of the major pheromonal sources in rat, the results suggest that α-2u globulin might act as a carrier for hydrophobic odorants of preputial gland.

Keywords: Pheromones, Preputial gland, Binding protein, Lipocalin, Rat, MALDI-TOF

*E-mail: garchu56@rediffmail.com

 

 

Indian Journal of Biochemistry & Biophysics

Vol. 43, October 2006, pp. 323-326

 

Extraction and carrier-facilitated transport of amino acids using synthetic non-cyclic receptors through bulk liquid membrane

 

Pratibha Joshi, Nidhi Joshi and Uma Sharma*

School of Studies in Chemistry, Vikram University,
Ujjain (MP) 456 010, India

Received 12 November 2005; revised 3 July 2006

The extraction and carrier-facilitated transport of amino acids (leucine, valine and glycine) was studied through chloroform bulk liquid membrane system using a series of non-cyclic receptors such as diethylene glycol (1), diethylene glycol dimethyl ether (2), diethylene glycol dibutyl ether (3), diethylene glycol dibenzoate (4), triethylene glycol (5) and tetraethylene glycol (6). The amount of amino acid extracted and transported depends mainly upon the structure and the concentration of the receptors and also on the concentration of amino acid. The receptors 1 to 4, having small chain length and flexible end groups, formed stable complexes with amino acids, and the flexibility of receptors in different conformational forms was responsible for their carrier ability, while the receptors 5 and 6, having larger chain length showed poor carrier ability. Hydrophobicity of amino acids also play an important role in the extraction as well as transport process.

Keywords:           Extraction, Liquid membrane, Carrier-facilitated transport, Receptors.

*E.mail: umasharma10@rediffmail.com

 

 

 

 

 

 

 

AUTHOR INDEX


Ali I A F

312

Mahmood A

267

Angayarkanni N

275

Mahmood S

267

Archunan G

319

Meng C X

284

Chatterjee M

299

Mulimani V H

295

Chi Z

289

Naseem I

312

Das S K

306

Ponmanickam P

319

Fatima R A

312

Qin S

284

Gao L

289

Ramakrishnan S

275

Girigowda K

295

Selvi R

275

Husain E

312

Sharma U

323

Joshi N

323

Sil P C

299

Joshi P

323

Su Z-L

284

Kaur K

267

Sulochana K N

275

Lakshmi S

275

Thippeswamy S

295

Liang L

289

Vasudevan D M

306

Ma L

289

Wei W

284


 

 

Award Announcement

 

Drs Anant and Lata Labhsetwar Charitable Trust Labhsetwar Foundation, USA

(E-mail: latalabh@aol.com)

 

Drs Anant and Lata Charitable Trust, Nagpur and Labhsetwar Foundation, USA have established a Population Control and Family Welfare Award, which is given annually to an individual of Indian nationality who has made outstanding research contributions in this field. The award consists of Rs 1,00,000 in cash, a memento and a certificate. The Committee has the right to split the award in the event found necessary.

The purpose of the award is to encourage and recognize the work of those who have made significant contributions towards India’s population stabilization issues. We recognize that work in various fields such as physiology, immunology, pharmacology, clinical, social, operational and behavioural work as well as demography is important in contributing towards population stabilization. Nominee should be under 55 years of age.

 

Nominations giving complete biodata of the candidate along with list of publications and reprints of the five best publications in the related field should be sent to:
“Dr. C P Puri, Director, National Institute for Research in Reproductive Health,
Jehangir Merwanji Street, Parel, Mumbai 400012”. Last date for receiving nominations is
1st November 2006. The award function will be
held in February 2007.