Iturin A
目录号 : GC65075
Iturin A是一种源自枯草芽孢杆菌和相关细菌的环状脂肽，对致病性酵母和真菌具有较强的抗真菌（antifungal）活性。
Cas No.:52229-90-0
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
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- Purity: >98.50%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
| Cell experiment [1]: | |
Cell lines | HepG2 cells |
Preparation Method | Cells were seeded in 96 wells plate at a density of 1×104 cells per well. After 4h, Iturin A was added to the cell culture at different working concentrations of 5μM, 10μM, 25μM, 50μM, 75μM, and 100μM, respectively. After 48h, cell viability was determined by MTT assay. |
Reaction Conditions | 5, 10, 25, 50, 75, 100μM; 48h |
Applications | Iturin A treatment inhibited the growth of HepG2 cells in a concentration-dependent manner. |
| Animal experiment [2]: | |
Animal models | Swiss albino mice |
Preparation Method | Sarcoma 180 cells were injected subcutaneously into the right flank of mice to generate solid tumors. After 78 days, tumors were grown and visible on the right flank of the mice. Tumor bearing mice were divided in different groups and injected Iturin A through tail vein. The treatment schedule was as following (1) Group1: Control (Sterile PBS), (2) Group2: 5mg/kg, (3) Group3: 10mg/kg and (4) Group4: 15mg/kg. Animals were treated for 28 days allowing one day interval. During the study period tumor volume of each animal were measured and recorded. After the scheduled treatment, all animals were sacrificed, and finally tumor volume and weight were measured. |
Dosage form | 5, 10, 15mg/kg for 28 days allowing one day interval; i.v. |
Applications | Iturin A induced apoptosis in sarcoma 180 tumors, as evidenced by decreased Bcl2, increased Bax expression, and cleavage of caspase and PARP. In addition, immunohistochemical analysis of tumor tissue sections revealed that CD 31, Ki67, pAkt, and pMAPK were down regulated in the Iturin A treated group. |
References: | |
Iturin A is a cyclic lipopeptide derived from Bacillus subtilis and related bacteria, which has strong antifungal activity against pathogenic yeast and fungi[1]. Iturin A can interact with ergosterol in the cytoplasmic membrane, leading to the formation of ion conduction pores and the outflow of intracellular ions, such as K+[2]. Iturin A has antiviral and antitumor activities[3].
In vitro, Iturin A (5-100μM) treatment of HepG2 cells for 48h inhibited cell growth in a concentration-dependent manner, significantly increased the proportion of cells in the G2/M phase, reduced the proportion of cells in the S phase, induced cell apoptosis, and led to an increase in intracellular ROS[4]. Iturin A (0-20μM) treated breast cancer MDA-MB-231 and MCF-7 cells for 48h significantly inhibited cell proliferation, with IC50 values of 7.98±0.19μM and 12.16±0.24μM, respectively. It induced cell apoptosis, changed the expression profile of proteins involved in cell apoptosis, and inhibited Akt kinase activity in cells[5]. Iturin A (0-100μM) treated Caco-2 cells for 48h inhibited cell growth in a concentration-dependent manner, induced cell apoptosis, upregulated the expression of apoptotic gene bax, and downregulated the expression of anti-apoptotic gene bcl-2[6].
In vivo, Iturin A (5, 10, 15mg/kg) was injected into the tail vein of mice bearing sarcoma 180 cells for 28 days, which significantly inhibited tumor growth, induced tumor cell apoptosis, upregulated Bax expression, increased caspase and PARP cleavage, and downregulated the expression of CD31, Ki67, pAkt, and pMAPK[7].
References:
[1] Yaraguppi D A, Bagewadi Z K, Patil N R, et al. Iturin: a promising cyclic lipopeptide with diverse applications[J]. Biomolecules, 2023, 13(10): 1515.
[2] Wang J, Qiu J, Yang X, et al. Identification of Lipopeptide Iturin A Produced by Bacillus amyloliquefaciens NCPSJ7 and Its Antifungal Activities against Fusarium oxysporum f. sp. niveum[J]. Foods, 2022, 11(19): 2996.
[3] Zhao H, Zhao X, Lei S, et al. Effect of cell culture models on the evaluation of anticancer activity and mechanism analysis of the potential bioactive compound, iturin A, produced by Bacillus subtilis[J]. Food & function, 2019, 10(3): 1478-1489.
[4] Zhao H, Yan L, Guo L, et al. Effects of Bacillus subtilis iturin A on HepG2 cells in vitro and vivo[J]. AMB Express, 2021, 11(1): 67.
[5] Dey G, Bharti R, Dhanarajan G, et al. Marine lipopeptide Iturin A inhibits Akt mediated GSK3β and FoxO3a signaling and triggers apoptosis in breast cancer[J]. Scientific reports, 2015, 5(1): 10316.
[6] Zhao H, Xu X, Lei S, et al. Iturin A‐like lipopeptides from Bacillus subtilis trigger apoptosis, paraptosis, and autophagy in Caco‐2 cells[J]. Journal of Cellular Physiology, 2019, 234(5): 6414-6427.
[7] Dey G, Bharti R, Banerjee I, et al. Pre-clinical risk assessment and therapeutic potential of antitumor lipopeptide ‘Iturin A’in an in vivo and in vitro model[J]. RSC advances, 2016, 6(75): 71612-71623.
Iturin A是一种源自枯草芽孢杆菌和相关细菌的环状脂肽，对致病性酵母和真菌具有较强的抗真菌（antifungal）活性[1]。Iturin A能够与细胞质膜中的麦角甾醇相互作用，导致离子传导孔的形成和细胞内离子的流出，例如K+[2]。Iturin A具有抗病毒、抗肿瘤活性[3]。
在体外，Iturin A（5-100μM）处理HepG2细胞48h，以浓度依赖性的方式抑制了细胞的生长，显著增加了G2/M期细胞比例，减少了S期细胞比例，诱导了细胞凋亡，导致了细胞内ROS增加[4]。Iturin A（0-20μM）处理乳腺癌MDA-MB-231和MCF-7细胞48h，显著抑制了细胞增殖，IC50 值分别7.98±0.19μM、12.16±0.24μM，诱导了细胞凋亡，改变了参与细胞凋亡的蛋白质的表达谱，抑制了细胞中的Akt激酶活性[5]。Iturin A（0-100μM）处理Caco-2细胞48h，以浓度依赖性的方式抑制了细胞的生长，诱导了细胞凋亡，上调了凋亡基因bax的表达，下调了抗凋亡基因bcl-2的表达[6]。
在体内，Iturin A（5, 10, 15mg/kg）通过尾静脉注射治疗肉瘤180细胞异种移植小鼠28天，显著抑制了肿瘤生长，诱导了肿瘤细胞凋亡，上调了Bax的表达，增加了caspase和PARP裂解，下调了CD31、Ki67、pAkt和pMAPK的表达[7]。
| Cas No. | 52229-90-0 | SDF | Download SDF |
| 分子式 | C48H74N12O14 | 分子量 | 1043.17 |
| 溶解度 | Ethanol : 5 mg/mL (4.79 mM; Need ultrasonic) H2O : 1 mg/mL (0.96 mM; Need ultrasonic) | 储存条件 | Store at -20°C |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 0.9586 mL | 4.7931 mL | 9.5862 mL |
| 5 mM | 0.1917 mL | 0.9586 mL | 1.9172 mL |
| 10 mM | 0.0959 mL | 0.4793 mL | 0.9586 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Iturin A Rescued STb-R-Induced Pork Skeletal Muscle Growth Restriction through the Hypothalamic-Pituitary-mTORC1 Growth Axis
Animals (Basel) 2022 Jun 17;12(12):1568.PMID:35739903DOI:10.3390/ani12121568.
The engineered STb-Rosetta Escherichia coli (STb-R) was designed to investigate the effects of Iturin A on the skeletal muscle growth of weaned piglets. A total of 28 piglets were randomly divided into 4 groups (7 piglets per group): the control group (100 mL PBS), the Iturin A group (100 mL 320 mg/kg body weight (BW) Iturin A), the STb-R group (100 mL 1 × 1010 CFU/mL STb-R), and the Iturin A + STb-R group (100 mL 320 mg/kg BW Iturin A + 1 × 1010 CFU/mL STb-R). Compared with the control, STb-R-reduced body weight gain were rescued by Iturin A. The semimembranosus muscle weight recovered to normal level in the Iturin A + STb-R group. The level of relevant genes of the growth axis were elevated by Iturin A, including GHRH in the hypothalamus, GHRHR and GH in the pituitary, and GHR, IGF-1 and IGF-1R in the semimembranosus muscle. Moreover, Iturin A increased the mean fiber area and the number of proliferating cells in the semimembranosus muscle, which were decreased by STb-R. Additionally, the mTORC1 pathway was reactivated by Iturin A to relieve the suppression of STb-R. Collectively, the hypothalamic-pituitary growth axis-mediated Iturin A reactivated the mTORC1 pathway to rescue STb-R-restricted pork skeletal muscle growth.
Enhanced production of Iturin A by strengthening fatty acid synthesis modules in Bacillus amyloliquefaciens
Front Bioeng Biotechnol 2022 Sep 9;10:974460.PMID:36159706DOI:10.3389/fbioe.2022.974460.
Iturin A is a biosurfactant with various applications, and its low synthesis capability limits its production and application development. Fatty acids play a critical role in cellular metabolism and target product syntheses, and the relationship between fatty acid supplies and Iturin A synthesis is unclear. In this study, we attempted to increase Iturin A production via strengthening fatty acid synthesis pathways in Bacillus amyloliquefaciens. First, acetyl-CoA carboxylase AccAD and ACP S-malonyltransferase fabD were overexpressed via promoter replacement, and Iturin A yield was increased to 1.36 g/L by 2.78-fold in the resultant strain HZ-ADF1. Then, soluble acyl-ACP thioesterase derived from Escherichia coli showed the best performance for Iturin A synthesis, as compared to those derived from B. amyloliquefaciens and Corynebacterium glutamicum, the introduction of which in HZ-ADF1 further led to a 57.35% increase of Iturin A yield, reaching 2.14 g/L. Finally, long-chain fatty acid-CoA ligase LcfA was overexpressed in HZ-ADFT to attain the final strain HZ-ADFTL2, and Iturin A yield reached 2.96 g/L, increasing by 6.59-fold, and the contents of fatty acids were enhanced significantly in HZ-ADFTL2, as compared to the original strain HZ-12. Taken together, our results implied that strengthening fatty acid supplies was an efficient approach for Iturin A production, and this research provided a promising strain for industrial production of Iturin A.
Effects of Bacillus subtilis Iturin A on HepG2 cells in vitro and vivo
AMB Express 2021 May 10;11(1):67.PMID:33970365DOI:10.1186/s13568-021-01226-4.
Iturin A with cyclic peptide and fatty acid chain isolated from Bacillus subtilis fermentation shows a variety of biological activities. Among them, the anticancer activity attracted much attention. However, the molecular mechanism of its inhibitory effect on hepatocellular carcinoma was still unclear. Thus its effect on hepatocellular carcinoma was tested in this research. It was found that Iturin A could enter HepG2 cells immediately and cause reactive oxygen species burst, disrupt cell cycle and induce apoptosis, paraptosis and autophagy in vitro. The Iturin A without fatty acid chain showed no antitumor activity. Amphiphilic is critical to the activity of Iturin A. The anticancer activity of Iturin A to hepatocellular carcinoma was also verified in mice models carrying xenograft tumors constructed by HepG2 cells. At a dosage of 3 mg/kg/day, Iturin A significantly inhibited the further increase of the tumor weight by 58.55%, and reduced the expression of Ki67 in tumor. In the tumor treated with Iturin A, lymphocyte infiltration was found, and the expressions of TGF-β1and PD-L1 were decreased, which indicated that the tumor immune microenvironment was improved. Besides, Iturin A showed no significant harm on the health of mice except slight disturbance of liver function. These results suggested that Iturin A had significant antitumor effect in vitro and vivo, and provide a basis for the application of Iturin A as anticancer agent.
Gene Expression and Characterization of Iturin A Lipopeptide Biosurfactant from Bacillus aryabhattai for Enhanced Oil Recovery
Gels 2022 Jun 25;8(7):403.PMID:35877488DOI:10.3390/gels8070403.
Biosurfactants are eco-friendly surface-active molecules recommended for enhanced oil recovery techniques. In the present study, a potential lipopeptide (biosurfactant) encoding the Iturin A gene was synthesized from Bacillus aryabhattai. To improvise the yield of the lipopeptide for specific applications, current research tends toward engineering and expressing recombinant peptides. An Iturin A gene sequence was codon-optimized, amplified with gene-specific primers, and ligated into the pET-32A expression vector to achieve high-level protein expression. The plasmid construct was transformed into an E. coli BL21 DE3 host to evaluate the expression. The highly expressed recombinant Iturin A lipopeptide was purified on a nickel nitrilotriacetic acid (Ni-NTA) agarose column. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) revealed that the purity and molecular mass of Iturin A was 41 kDa. The yield of recombinant Iturin A was found to be 60 g/L with a 6.7-fold increase in comparison with our previously published study on the wild strain. The approach of cloning a functional fragment of partial Iturin A resulted in the increased production of the lipopeptide. When motor oil was used, recombinant protein Iturin A revealed a biosurfactant property with a 74 ± 1.9% emulsification index (E24). Purified recombinant protein Iturin A was characterized by mass spectrometry. MALDI-TOF spectra of trypsin digestion (protein/trypsin of 50:1 and 25:1) showed desired digested mass peaks for the protein, further confirming the identity of Iturin A. The Iturin A structure was elucidated based on distinctive spectral bands in Raman spectra, which revealed the presence of a peptide backbone and lipid. Recombinant Iturin A was employed for enhanced oil recovery through a sand-packed column that yielded 61.18 ± 0.85% additional oil. Hence, the novel approach of the high-level expression of Iturin A (lipopeptide) as a promising biosurfactant employed for oil recovery from Bacillus aryabhattai is not much reported. Thus, recombinant Iturin A demonstrated its promising ability for efficient oil recovery, finding specific applications in petroleum industries.
Iturin A Induces Resistance and Improves the Quality and Safety of Harvested Cherry Tomato
Molecules 2021 Nov 16;26(22):6905.PMID:34833997DOI:10.3390/molecules26226905.
The soft rot disease caused by Rhizopus stolonifer is an important disease in cherry tomato fruit. In this study, the effect of Iturin A on soft rot of cherry tomato and its influence on the storage quality of cherry tomato fruit were investigated. The results showed that 512 μg/mL of Iturin A could effectively inhibit the incidence of soft rot of cherry tomato fruit. It was found that Iturin A could induce the activity of resistance-related enzymes including phenylalanine ammonia lyase (PAL), polyphenol oxidase (PPO), peroxidase (POD), glucanase (GLU), and chitinase (CHI), and active oxygen-related enzymes including ascorbate peroxidases (APX), superoxide dismutases (SOD), catalases (CAT), and glutathione reductase (GR) of cherry tomato fruit. In addition, Iturin A treatment could slow down the weight loss of cherry tomato and soften the fruit. These results indicated that Iturin A could retard the decay and improve the quality of cherry tomato fruit by both the inhibition growth of R. stolonifera and the inducing the resistance.