In the described study , the bio-enhancement of Rosmarinic acid( RA) was done successfully in suspension culture of Salvia officinalis and Ocimum sanctum suspended with E coli K12 entrapped alginate beads.It is reported that Tyrosin Amino transferase has a key role in the production of Rosmarinic acid , which is present in E coli K 12. Quantification by HPTLC indicated that the maximum RA content was noted in 4 weeks old (2050 mg/l) in Salvia officinalis suspension culture fed with Tyrosin 10 mg/100ml followed by 4 weeks old Ocimum sanctum suspension culture fed with Tyrosin 10 mg/100ml(715 mg/l), that are about 18 fold and 5 folds enhancement as compared to control. The method for significantly enhancement of RA was reported for the first time.
Table of Contents
1. Introduction
2. Material and Methods
2.1 Plant material
2.2 Suspension media
2.3 Preparation of beads of E coli K12
2.4 Down stream process for Rosmarinic acid extraction
2.5 High Performance Thin Layer Chromatography (HPTLC) Analysis
2.6 Infra Red Spectral Studies
2.7 Statistical analysis
3. Results and Discussion
4. Conclusion
Research Objectives and Themes
The primary objective of this research is to enhance the production of Rosmarinic acid, a medicinally valuable secondary metabolite, in Salvia officinalis and Ocimum sanctum suspension cultures. The study investigates a novel biotechnological approach using immobilized E. coli K12 cells in alginate beads as elicitors to stimulate the tyrosine metabolic pathway, aiming to establish an economically viable and scalable industrial production method.
- Bio-enhancement of secondary metabolites through in vitro cell culture technology.
- Implementation of immobilized E. coli K12 alginate beads as novel biotic elicitors.
- Optimization of tyrosine-mediated biosynthesis of Rosmarinic acid.
- Quantification and characterization of metabolite yield using HPTLC and FTIR spectral analysis.
Excerpt from the Book
Results and Discussion
Plant Tissue culture has been served as a mean for secondary metabolites production. The strong and growing demand in today's marketplace has refocused attention on in vitro plant materials as potential factories for secondary phytochemical products. Chaturvedi et al., 2009, 2010, 2011, 2013, 2014, 2015 , 2016 have extensively studied the enhancement of secondary metabolites by using different compound in tissue culture of medicinal plants. The treatment of plant cells with biotic and/or abiotic elicitors has been a useful strategy to enhance secondary metabolite production in cell cultures( Karuppusamy et al., 2009). Tyr B gene of E. coli strain K-12 govern the production of Tyrosin Amino Transferase (TAT) . TAT, a broad-specificity enzyme, which catalyzes the transamination of 2-ketoisocaproate, p-hydroxy phenyl pyruvate, and phenyl pyruvate to yield leucine, tyrosine and phenylalanine, respectively . (Huang, 2009).
Hence keeping all the facts in view,in the present study various elicitors were used to bio-enhance the Rosmarinic acid(RA)by using tissue culture technique. The production of RA, a caffiec acid ester compound of medicinal value has been successfully induced by using alginate beads of E coli K12 bacterium in suspension culture of Salvia officinalis and Ocimum sanctum separately supplemented with various elicitors. Enhancement as observed in all the samples but the maximum content was noted in 4 weeks old (2050 mg/l) in Salvia officinalis suspension culture fed with Tyrosin 10 mg/100ml followed by 4 weeks old Ocimum sanctum suspension culture fed with Tyrosin 10 mg/100ml(715 mg/l), that are about 18 fold and 5 folds enhancement as compared to control (Table 1,2)..It is already reported. That this extra cellular enzyme catalyzes the first step of the tyrosine pathway leading to the formation of rosmarinic acid (alpha-O-caffeoyl-3,4-dihydroxyphenyllactic acid) in suspension cultures of Anchusa officinalis L(De-Eknamkul et al., 1887) .This study supports the enhanced production of Rosmarinic acid in our present findings.
Summary of Chapters
Introduction: Provides the biological and medicinal background of Rosmarinic acid and explains the rationale for using E. coli K12 as a source of Tyrosine aminotransferase for biosynthetic enhancement.
Material and Methods: Details the experimental setup, including plant material preparation, suspension media composition, the protocol for creating immobilized E. coli beads, and the analytical techniques (HPTLC/FTIR) used.
Results and Discussion: Presents the quantitative data on Rosmarinic acid production, confirming significant fold-increases in treated cultures compared to controls and discussing the role of enzymatic stress in the pathway.
Conclusion: Summarizes the success of the methodology in achieving high yields and emphasizes the potential for this technique in industrial-scale production.
Keywords
Rosmarinic acid, Salvia officinalis, Ocimum sanctum, suspension culture, E. coli K 12, Tyrosin Amino Transferase, HPTLC, FTIR, secondary metabolites, bio-enhancement, cell culture technology, plant biotechnology, medicinal plants.
Frequently Asked Questions
What is the core focus of this research?
The research focuses on increasing the yield of Rosmarinic acid in medicinal plant cell cultures using a novel bio-engineering approach with immobilized bacteria.
What are the primary subjects of the study?
The study examines Salvia officinalis and Ocimum sanctum, both recognized for their secondary metabolite content.
What is the main goal of the project?
The primary goal is to achieve significant enhancement in the production of Rosmarinic acid, a compound with high medicinal value, using cost-effective biotechnological methods.
Which scientific methods were employed?
The methodology includes plant tissue culture, enzymatic elicitation via immobilized E. coli K12, High Performance Thin Layer Chromatography (HPTLC) for quantification, and FTIR for spectral characterization.
What is discussed in the main part of the paper?
The main section covers the preparation of media and biological elicitors, the experimental procedure of suspension culture, and a detailed analysis of the biochemical results obtained.
Which keywords characterize this work?
Key terms include Rosmarinic acid, bio-enhancement, suspension culture, E. coli K12, and HPTLC analysis.
Why was E. coli K12 specifically used in this study?
E. coli K12 was selected because it contains the Tyrosine aminotransferase enzyme, which is crucial for the biosynthesis of Rosmarinic acid.
What were the most significant quantitative results?
The study observed an 18-fold increase in Rosmarinic acid content in Salvia officinalis and a 5-fold increase in Ocimum sanctum after four weeks of treatment.
How does the use of alginate beads contribute to the process?
Alginate beads are used for the immobilization of E. coli cells, which subjects the bacteria to stress, thereby enhancing the production and extracellular release of the target enzyme.
- Citar trabajo
- Dr. Pratibha Chaturvedi (Autor), 2017, Microbe supported enhanced production of Rosmarinic acid of medicinal plants in vitro, Múnich, GRIN Verlag, https://www.grin.com/document/355224
