商品描述

Plant Materials:

Soybean plants (Glycine max cv. Jutro) were grown in nitrogen-free medium in a growth chamber at 26°C under a photoperiod of 16 h. Inoculation of plants with Bradyrhizobium japonicum USDA 110 was performed directly upon sowing, and nodules were collected 4 wk after inoculation. Uninfected soybean plants were cultured in the same way. Nodules and uninfected roots were frozen in liquid nitrogen immediately after harvesting and stored at 70°C.

Construction of Plasmids:

The 0.68-kb fragment of Glycine max ENOD40 cDNA (European Molecular Biology Laboratory database, accession no. X69154; ref. 4) was cloned into the BamHI and SalI sites of pBluescript SK (Stratagene). Single point mutations in the ATG of ORF A or ORF B were introduced into the ENOD40 gene by using the QuikChange site-directed mutagenesis kit (Stratagene). The mutations were confirmed by DNA sequencing.

Synthetic Peptides:

Peptides were synthesized by Neosystem (Strasbourg, France) according to the amino acid sequence deduced from the nucleotide sequence of ORF A (MELCWLTTIHGS) and ORF B (MVLEEAWRERGVRGEGAHSSHSLT) of soybean ENOD40 cDNA. Synthetic analogs were MELSWLTTIHGS and MELCWLTTIGGG for peptide A and EVGHSRAWHASEGLRMTSRLEGVE and MVLQQAWGQGGVGGQGAYSSYSLT for peptide B. Peptides were labeled by coupling an additional biotinylated lysine residue to the C terminus of the sequence. Control peptides were from Bachem and biocytin (biotinyl-L-lysin) was from Sigma. Control A was a biotinylated analog of the cGMP-dependent protein kinase substrate with the sequence RKISASEFDRPLR. Control B was a biotinylated fragment (residues 44-68) of human PTH (RDAGSQRPRKKEDNVLVESHEKSLG). Purity of each peptide was assessed by HPLC.

Peptide-Binding Assays:

Peptide-binding assays were performed with partially purified sucrose synthase (SuSy). Soybean nodules were homogenized with extraction buffer containing 5 mM DTT as described above. After centrifugation at 30,000 × g at 4°C, the supernatant (500 µl) was fractionated on a calibrated Superdex 200 HR 10/30 column (Amersham Pharmacia). The column was eluted with 20 mM Tris·HCl (pH 8.0), 10 mM MgCl2, and 200 mM NaCl (for binding assays with peptide A) or with 20 mM Tris·HCl (pH 8.0), 10 mM MgCl2, and 500 mM NaCl (for assays with peptide B) at a flow rate of 0.8 ml/min. The SuSy-containing fraction ( 1.6 ml) was collected, and protease inhibitors and a detergent mix were added to achieve a final concentration of 0.1% Nonidet P-40 and 0.05% deoxycholate. Binding reactions with 5 µg of biotinylated peptides and subsequent coupling to streptavidin-agarose beads were performed as described above. The beads were washed five times with 20 mM Tris·HCl (pH 8.0), 10 mM MgCl2, 0.1% Nonidet P-40, and 0.05% deoxycholate containing 0.5 M NaCl (for peptide A-affinity matrix) or with the same buffer containing 1 M NaCl (for peptide B-affinity matrix). Before SDS/10% PAGE, the beads were washed once with 10 mM Tris·HCl (pH 8.0), 5 mM MgCl2, and 50 mM NaCl.

The product contains Soybean Peptide which is extracted from the Soy Protein Isolate through enzyme reaction, is composed of 3 to 6 amino acid with molecular weight less than 1,000d. Peptide is absorbed and digested by our body more directly than amino acid, is low in sensitivity and zero in cholesterol that is good to health. It also consists of complete plant protein, rich in 9 kinds of essential amino acids, that make rapid recovery of energy, accelerate the development of brain power and enhance memory. Soybean peptide is also good for energy metabolism, helping to keep a healthy body shape.

Different extraction techniques obtain different kinds of peptide.SoyPeptide Powder - Anti-fatik Formula keeps most of the major ingredients of soybean including Saponin, Isoflavone, Unsaturated Fatty Acid, Minerals, Vitamins, Lecithin and Soybean Oligose. The product contains no stimulant as tested by the Analeptic Inspection Center of Athletic Iatrology Institute of State Sports General Bureau. It is particularly suitable for people with problems of progressive fatigue, poor quality of sleeping, irregular working schedules and overstrained mind.

Soybean Peptide is extracted from the Soy Protein Isolate through enzyme reaction, which is composed of 3 to 6 amino acid with molecular weight less than 1,000d.

20 kinds of amino acid (arginine, glutamic acid, serin, aspartic acid, leucine, etc.) are changed to oligopeptide(known as low-molecular-weight substance). Therfore, it is easy to absorb into the body.

Antimitotic and Cancer Preventive Properties of a Soybean Peptide:

Ben O. de Lumen and Alfredo F. Galvez; Nutritional Sciences and Toxicology, University of California at Berkeley, Berkeley, CA, USA. . Third International Symposium on the Role of Soy in Preventing and Treating Chronic Disease. October 31 - November 3, 1999 Omni Shoreham Hotel Washington, DC USA

Epidemiologic evidence suggesting the correlation between diets high in soybean and overall low cancer mortality rates, especially those of colon, breast, and prostate, has given impetus to identifying components in soybean responsible for its anticancer properties.

We isolated a soybean cDNA encoding the small subunit peptide of a cotyledon-specific 2S albumin (Gm2S-1). The peptide (named lunasin) has a unique, highly acidic carboxyl end. A chimeric gene encoding the lunasin peptide tagged with green fluorescent protein arrested cell division and caused abnormal spindle fiber elongation, chromosomal fragmentation, and cell lysis when transiently transfected into murine embryo fibroblast, murine hepatoma, and human breast cancer cells. Deletion of the acidic carboxyl end abolished the antimitotic effect. Immunolocalization of lunasin and an immuno-binding assay using synthetic peptides revealed the preferential adherence of lunasin to chromatin.

Lunasin is the first antimitotic peptide whose cDNA was cloned and the first from a common food source. We also showed that the exogenous application of lunasin peptide to as low as 125 nmol/L inhibits the in vitro transformation of mouse embryo fibroblast cells (C3H 10T 1/2) into tumorous foci by the carcinogens 3-methyl cholanthrene and 7,12-dimethylbez[a]anthracene.

The lunasin gene has potential application as an antimitotic cancer therapeutic agent, and the lunasin peptide may be an important cancer-preventive compound in soybeans. Furthermore, its antimitotic property suggests that lunasin could play a native role in arresting mitosis that initiates the cell expansion phase of seed development where DNA endoreduplication and synthesis of storage proteins, lipids, and carbohydrates occur.

In the last few months another energy source entered the spotlight -soybean peptide:

Soy is the most widely used botanical for foods and supplements such as pre- and post-menopausal women, but it is used for many other applications including lowering cholesterol and in sports beverages. Soybean peptide (SP) is a bonding of multiple amino acids and it absorbs in the intestinal tract faster and more efficiently than amino acids and it seems to help the body regain lost energy quickly and may produce energy faster than other amino acids. SP has been shown to take only 20 minutes to absorb in the body, so those wanting to regain lost energy will take soy peptide within 30 minutes after the energy loss or one hour before sleeping. SP has also been studied for diet, enhancement of brain function and beautifying the skin.

A soy ingredient used extensively in supplements could hold the power to inhibit the growth of tumour cells, says Chinese biotech firm American Oriental Bioengineering, announcing new findings for soybean protein peptides:

The company’s research team reports that the growth of liver cancer cells inside mice was ‘successfully suppressed’ by soybean protein peptides without side effects. Their findings could lead to future market opportunities for the ingredient.

Soy is the most widely used botanical by pre- and post-menopausal women but its use by men is also growing as research continues to show the benefits on heart health. In the US, soy sales have grown from $940 million in 1990 to a projected $4 billion this year.

The World Health Organisation (WHO) estimates that dietary factors are estimated to account for approximately 30 per cent of cancers in western countries, making diet second only to tobacco as a preventable cause of cancer.

Overweight and obesity are both serious risk factors for cancer. Diets high in fruit and vegetables may reduce the risk for various types of cancer, while high levels of preserved and/or red meat consumption are associated with increased cancer risk. More than 7 million people now die each year from cancer.

Research conducted at American Oriental Bioengineering on the impact of soy protein peptides focused on stopping the growth of the metastatic tumour cells because of their ability to mutate more quickly than normal cells, which gives them a greater ability to adapt to their environment, as well as a greater ability to resist therapy.

"Because the liver is close to or actually connected to several significant organs, and because the liver plays an important role in blood circulation by acting as a filter, metastatic liver cancer occurs in over 75 per cent of all terminal cancer patients," said the firm.

The company's research team discovered that soybean protein peptide extracted from particular species of soybean could effectively inhibit the growth of tumours.

Diverse soybean accessions from the U.S. Department of Agriculture Soybean Germplasm Collection, several commercially available soy protein fractions and isoflavone-enriched products. With synthetic lunasin and monoclonal antibody, ELISA shows a linear concentration range of 24-72 ng/mL, good reproducibility, a detection limit of 8 ng/mL, and a recovery of 90% on spiked soy samples. Lunasin concentrations in the tested materials range from 0.10 to 1.33 g/100 g flour. Differences that exceeded 100% have been observed among accessions of similar maturity that were grown in the same environment, indicating that genetic differences in soybeans exist for lunasin. Soy protein concentrate, isolate, and hydrolyzate contain 2.8 , 3.75, and 4.4 g lunasin/100 g flour, respectively, while soy flour and soy flakes contain 1.24 g lunasin/100 g flour. Isoflavone-enriched products contain very little or no lunasin. The wide range of lunasin concentrations within the Glycine max species indicates that the levels of this important bioactive peptide can be genetically manipulated. Furthermore, soy isolates and hydrolyzed soy proteins contain the highest concentrations of lunasin.

Lunasin suppresses E1A-mediated transformation of mammalian cells but does not inhibit growth of immortalized and established cancer cell lines:

Lunasin, a novel and promising chemopreventive compound isolated from soybean cotyledon, is a 43-amino acid peptide that contains a -RGD-cell adhesion motif followed by 8 aspartic acid residues at the carboxyl end and a structurally conserved helix region. We showed previously that lunasin peptide applied exogenously reduces foci formation in mouse fibroblast cells treated with chemical carcinogens and inhibits skin tumorigenesis induced by chemical carcinogens in mice when applied topically. In this study, lunasin peptide applied to cell culture suppresses foci formation in E1A-transfected mouse fibroblast NIH 3T3 cells. Within 18 h of exogenous application, lunasin internalizes into the cell and localizes in the nucleus. In an initial study of genes affected by lunasin, the peptide increases p21 protein levels fivefold in cells transfected with E1A but not in untransfected cells. In contrast to its inhibitory effects on cell transformation, lunasin has no effect on growth of imicroMortalized (nontumorigenc) and established cancer cells. This is the first report that lunasin suppresses transformation of mamicroMalian cells induced by an oncogene (E1A) in addition to chemical carcinogens.

Characterization of lunasin isolated from soybean:

Lunasin is a novel and promising chemopreventive peptide from soybean. We have shown previously that lunasin suppresses transformation of mammalian cells caused by chemical carcinogens and inhibits skin carcinogenesis in mice when applied topically. Although the lunasin gene was cloned from soybean, all experiments carried out so far in our lab have used synthetic lunasin and therefore there is no detailed description of natural lunasin isolated from soybean. We report here the first characterization of soybean lunasin that includes definitive identification by mass peptide mapping, partial purification, and measurement of bioactivities of the various purified fractions and protein expression in the developing seed. The identity of lunasin in the seed extracts was established by Western blot analysis and mass spectrometric peptide mapping. All lunasin fractions partially purified by anion exchange and immunoaffinity column chromatography suppress colony formation induced by the ras-oncogene and inhibit core H3-histone acetylation. During seed development, lunasin peptide appears 5 weeks after flowering and persists in the mature seed. These results demonstrate the feasibility of producing large quantities of natural lunasin from soybean for animal and human studies.

The anticarcinogenic potential of soybean lectin and lunasin:

Cancer is one of the leading causes of death worldwide, generally exceeded only by cardiovascular disease in the developed world. The number of people diagnosed with cancer within the next few decades is expected to double. There will therefore be increased demand for novel diagnostic and medical therapies that use new non-traditional sources. Soybeans contain a variety of anticarcinogenic phytochemicals. Recently, there has been increased interest in the potential health benefits of bioactive polypeptides and proteins from soybeans, including lunasin and lectins. Lunasin is a polypeptide that arrests cell division and induces apoptosis in malignant cells. Lectins are glycoproteins that selectively bind carbohydrates; lectins are used in medicine in a variety of new applications. Additional research, including clinical trials, should continue to examine and elucidate the therapeutic effects, nutritional benefits, and toxic consequences of commonly ingested soybean lectins and lunasin.

Barley lunasin suppresses ras-induced colony formation and inhibits core histone acetylation in mammalian cells:

Lunasin is a novel peptide originally identified in soybean that suppresses chemical carcinogen-induced transformation in mammalian cells and skin carcinogenesis in mice. Since the lunasin gene was cloned from soybean and the chemically synthesized form of the lunasin peptide has been used in experiments conducted so far, the isolation of lunasin from other natural sources and testing of its biological properties have not been carried out. We report here the isolation, purification, and biological assay of lunasin from barley, a newly found rich source of the peptide. The identity of lunasin was established by Western blot analysis and mass spectrometric peptide mapping of the in-gel tryptic digest of the putative protein band. Lunasin was partially purified with anion exchange and immunoaffinity chromatography. The crude and partially purified lunasin from barley suppressed colony formation in stably ras-transfected mouse fibroblast cells induced with IPTG. These fractions also inhibited histone acetylation in mouse fibroblast NIH 3T3 and human breast MCF-7 cells in the presence of the histone deacetylase inhibitor sodium butyrate.

Chemopreventive property of a soybean peptide (lunasin) that binds to deacetylated histones and inhibits acetylation:

Lunasin is a unique 43-amino acid soybean peptide that contains at its carboxyl end: (a) nine Asp (D) residues; (b) an Arg-Gly-Asp (RGD) cell adhesion motif; and (c) a predicted helix with structural homology to a conserved region of chromatin-binding proteins. We demonstrated previously that transfection of mammalian cells with the lunasin gene arrests mitosis, leading to cell death. Here we show that exogenous application of the lunasin peptide inhibits chemical carcinogen-induced transformation of murine fibroblast cells to cancerous foci. To elucidate its mechanism of action we show that lunasin: (a) internalizes in the cell through the RGD cell adhesion motif; (b) colocalizes with hypoacetylated chromatin; (c) binds preferentially to deacetylated histone H4 in vitro; and (d) inhibits histone H3 and H4 acetylation in vivo in the presence of a histone deacetylase inhibitor. These results suggest a mechanism whereby lunasin selectively induces apoptosis, mostly in cells undergoing transformation, by preventing histone acetylation. In support of this, lunasin selectively induces apoptosis in E1A-transfected cells but not in nontransformed cells. Finally, in the SENCAR mouse skin cancer model, dermal application of lunasin (250 microg/week) reduces skin tumor incidence by approximately 70%, decreases tumor yield/mouse, and delays the appearance of tumors by 2 weeks relative to the positive control. These results point to the role of lunasin as a new chemopreventive agent that functions possibly via a chromatin modification mechanism.

Of all vegetable proteins, only the soy protein provides 2-5 year old children with all the essential amino acids specified by FAO/WHO/UNU in 1985. Amino acid composition of soybean peptide is almost the same as that of soy protein. Those babies allergic to milk protein can use it as a milk substitute.

Recent metabolic studies have shown that dietary proteins are mostly hydrolyzed into peptides by digestive enzyme in the stomach and intestine and then are digested and absorbed by the intestine in the form of peptides. Labeled isotope experiments in vivoshow that the soybean peptide has higher digestibility than milk protein and general amino acids do. Clinically, soybean peptide can be used as nutrition supplement for dyspepsia babies and seniors, convalescents after surgeries (especially alimentary tract surgeries), people with strain of lumbar muscles, dyspepsia persons requiring a large amount of proteins, and people antigenically allergic to proteins. It can help them return to balanced nutrition status sooner than injected amino acids.

Soybean peptide is an effective ingredient of α-glucosidase inhibitor. By slowing down the digestion of sucrose and starch, and preventing glucose level from rapidly rising, it can lower the glucose level.

Soybean peptide inhibits angiotensin inverting enzyme activity, so it can prevent peripheral blood vessel from contracting. Therefore, it can help hypertensive lower their blood pressure and has no effects on normal people. It can also reduce harmful LDL-cholesterol level in serum, but it does not decrease healthy HDL-cholesterol level. The more important is that soybean peptide can prevent cholesterol from rising after normal people take meat, eggs, and animal viscera with high cholesterol content. (The mechanism is unclear. The reason might be that soybean peptide can combine with cholesterol to prevent cholesterol absorption.)

Animal experiments in Japan showed after rats were fed with soybean peptide, the activity of heat generating fat BAT was stimulated. The more soybean peptide is fed, the higher activity is. It was found that obese children lost weight by having soybean peptide, because soybean peptide can decrease subcutaneous fat deposition, and increase fat burning and basal metabolism. Therefore, it is a safe and effective weight loss food (medicine) for obese children. With another benefit of glucose reduction, it can be taken for a long time to effectively control weight.

Soybean peptide can promote microorganism (Bifidobucterium) growth, stimulate metabolism, proliferate lactobacillus and fungi, and increase CO2 production of bread yeasts. Due to its good water absorption and moisture retention, soybean peptide can be used as an addictive during bread and cake baking to avoid dryness and extend their valid storage time. Soybean peptide has good foamability. Studies show that its foamability is as 4 times high as general proteins.

Due to illness and senescence, seniors can hardly take enough protein. Since soybean peptide has excellent digestibility, it is a protein supplement for the old and weak.

Soybean peptide can promote microorganism growth and metabolism, so it can be used in yogurt, cheese, vinegar, bread and soy sauce production to increase production efficiency and to improve the stability, nutrition, and flavor of products. Generally, protein cannot be dissolved in acid beverages. However, soybean peptide is easy to be dissolved in acid beverages and the solubility is not affected by PH. Soybean peptide is able to increase the concentrations of alanine and leucine in the blood to produce stable NAD, which can facilitate alcohol metabolism and then decrease alcohol concentration in the body. Therefore, soybean peptide has a benefit of alcohol removing and liver protection. In addition, athletes’ strength is proportional to the muscle mass. The more exercise, the more energy (protein and amino acid) is needed. Soybean peptide has a lower molecular weight. It is much easier to be digested than proteins and amino acid. It can also expedite the lipid metabolism process of transferring fat to energy so as to provide athletes with energy and to help them recover from fatigue.

Physical and chemical properties:

1. Physical properties:

This product is the white or/and yellowish powder and water-soluble.

2. Chemical properties:

3. Hygiene characteristics