Analytical Methods for the Detection and Purification of ε-Poly-L-lysine for Studying Biopolymer Synthetases, and Bioelectroanalysis Methods for Its Functional Evaluation

Katano, Hajime; Uematsu, Kohei; Maruyama, Chitose; Hamano, Yoshimitsu

doi:10.2116/analsci.30.17

Cited by 20 publications

(12 citation statements)

References 41 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Thus, chitosan oligosaccharides hydrochloride salts with n ≥ 4 could be separated and purified from a commercial product with n ≥ 2. Such solubility-based or precipitation/dissolution techniques have been applied to the separation and purification of other natural products, [6][7][8][9][10] and have proven more advantageous than conventional column separation techniques.…”

Section: Introductionmentioning

confidence: 99%

Solubility-based Separation and Purification of Long-Chain Chitin Oligosaccharides with an Organic-Water Mixed Solvent

et al. 2017

Self Cite

View full text Add to dashboard Cite

A simple and rapid method for separation and purification of chitin oligosaccharides, (GlcNAc)n, with n ≥ 5 is presented. A commercially available chitin oligosaccharides sample, consisting of (GlcNAc)n with n = 1 -7, was used as the starting material. Ten milligrams of the material was mixed with 100 μL of the 1 mol/L HCl. All the (GlcNAc)n species were dissolved in the aqueous medium. The aqueous solution was mixed with 900 μL of EtOH; the mixture was centrifuged, and the supernatant was removed to obtain a precipitate. The precipitate was found to consist mainly of (GlcNAc)n with n ≥ 5, indicating the significant difference in solubility between the short-chain (GlcNAc)n species with n ≤ 3 and the longer ones. By the repetition of the operations, a high purity long-chain (GlcNAc)n sample with n ≥ 5 could be prepared successfully. Since the long-chain (GlcNAc)n species are known to have excellent elicitor activity, this sample would be useful in the study of plant pathology, as well as chitin and chitosan chemistry.

show abstract

Section: Introductionmentioning

confidence: 99%

Solubility-based Separation and Purification of Long-Chain Chitin Oligosaccharides with an Organic-Water Mixed Solvent

et al. 2017

Self Cite

View full text Add to dashboard Cite

show abstract

“…In the first operation of the colorimetric method, 1 mL of aqueous solution containing ~0.8 mEq/L 12-molybdosilicate 9,10 was used instead of 0.1 mL of the 10 mEq/L Na3AR solution.…”

Section: Other Precipitate Reagentmentioning

confidence: 99%

Colorimetric Microtiter Plate Assay of Polycationic Aminoglycoside Antibiotics in Culture Broth Using Amaranth

et al. 2017

Self Cite

View full text Add to dashboard Cite

We present a colorimetric method for the detection of aminoglycoside antibiotics such as neomycin (NEO) using a reddish anionic dye, amaranth (AR 3-). Under acidic conditions, at which NEO exists in fully protonated form (NEOH 6+ ), the AR 3-anion associates with the NEOH 6+ cation to form a precipitate, NEOH(AR)2. The precipitate was soluble in a buffer solution of pH 8.5, yielding a reddish solution with an absorption maximum at around 520 nm. Tobramycin and gentamycin, which exist as pentavalent cations under acidic conditions, gave almost the same results. On the other hand, kanamycin, amikacin and streptomycin, which would exist as tri-and tetravalent cations, were not precipitated. Thus, the AR 3-anion could be considered to be an analytical reagent for specific aminoglycosides with polycationic functionality. However, since the precipitation reaction was considerably affected by other anions, a separation method using the tetraphenylborate anion was employed as a pretreatment. The separation method involves precipitating the polycationic aminoglycosides with the tetraphenylborate anion, washing the precipitate with acetonitrile, and re-precipitating the aminoglycosides as hydrochloride salts. Thus, the present method was applied to a microtiter plate assay of the products in an NEO-producing culture broth.

show abstract

“…The precipitation reaction with hydrophobic anions can be utilized in the determination and separation of εPL. In previous papers, 9,10 we have presented a method for the separation of εPL from culture broth using tetraphenylborate anion. The method is based on the precipitation of polycationic εPL with the anion and the re-precipitation as the hydrochloride salt.…”

Section: Introductionmentioning

confidence: 99%

“…3), indicating that εPL was precipitated quantitatively as the fully protonated εPL (εPLH n+ ) salt with the DPA -anion, εPLH(DPA)n. The acid-base equilibrium of εPL is described elsewhere. 1,9,10 In the aqueous solution at pH 6.2, εPL would exist in a polycationic form (εPLH m+ , m < n) by the protonation of some α-amino groups, according to the reaction:…”

mentioning

confidence: 99%

Separation and Purification of ε-Poly-l-lysine with Its Colorimetric Determination Using Dipicrylamine

et al. 2015

Self Cite

View full text Add to dashboard Cite

A separation method for a biopolymer, ε-poly-L-lysine (εPL), with its colorimetric determination using a yellow anionic dye, dipicrylamine anion (DPA -), is presented. The εPL-producing culture broth was mixed with a NaDPA solution to precipitate the εPL in polycationic form with the DPA -anion. The precipitate was dissolved into acetonitrile (AN). The AN solution was yellowish, and gave an absorption maximum at around 420 nm. Thus, εPL in the culture broth can be assayed colorimetrically a with multiwell microtiter plate. By the addition of bis(triphenylphosphoranylidene)ammonium chloride, the polycationic εPL was precipitated from the AN solution as the hydrochloride salt. On the other hand, the DPA -anion remained in the AN solution as the quaternary ammonium salt. Thus, εPL larger than tetramer can be separated and purified after the high-throughput screening of the synthetic enzyme.Keywords Separation, colorimetry, ε-poly-L-lysine, dipicrylamine, high-throughput screening

show abstract

Analytical Methods for the Detection and Purification of ε-Poly-L-lysine for Studying Biopolymer Synthetases, and Bioelectroanalysis Methods for Its Functional Evaluation

Cited by 20 publications

References 41 publications

Solubility-based Separation and Purification of Long-Chain Chitin Oligosaccharides with an Organic-Water Mixed Solvent

Solubility-based Separation and Purification of Long-Chain Chitin Oligosaccharides with an Organic-Water Mixed Solvent

Colorimetric Microtiter Plate Assay of Polycationic Aminoglycoside Antibiotics in Culture Broth Using Amaranth

Separation and Purification of ε-Poly-l-lysine with Its Colorimetric Determination Using Dipicrylamine

Contact Info

Product

Resources

About