Deproteinized natural rubber latex (DPNR-latex) was treated with lipase and phosphatase in order to analyze the structure of the chain-end group (alpha-terminal). The enzymatic treatment decreased the content of long-chain fatty acid ester groups in DPNR from about 6 to 2 mol per rubber molecule. The molecular weight and intrinsic viscosity were reduced to about one-third after treatment with lipase and phosphatase. The Huggins' k' constant of the enzyme-treated DPNR showed the formation of linear rubber molecules. The molecular weight distribution of DPNR changed apparently after treatment with lipase and phosphatase. (1)H NMR spectrum of rubber obtained from DPNR-latex showed small signals due to monophosphate, di-phosphate and phospholipids at the alpha-terminus. Treatment of DPNR-latex with lipase and phosphatase decreased the relative intensity of the (1)H NMR signals corresponding to phospholipids, whereas no change was observed for the signals due to mono- and diphosphates. The residual mono- and diphosphate signals as well as some phospholipid signals after lipase and phosphatase treatments indicate that mono- and diphosphate groups are directly linked at the alpha-terminus with the modified structure, expected by aggregation or linking with phospholipid molecules.

The treatment of deproteinized natural rubber (DPNR) latex with phospholipases A(2), B, C, and D decreased significantly the long-chain fatty acid ester contents in DPNR and also the molecular weight and Higgins' k' constant, except for phospholipase D treatment. This indicates the presence of phospholipid molecules in NR, which combine rubber molecules together. Transesterification of DPNR resulted in the decomposition of the functional group at the terminal chain-end (alpha-terminal), including phospholipids and formed linear rubber molecules. The addition of small amounts of ethanol into the DPNR solution reduced the molecular weight and shifted the molecular weight distribution (MWD) comparable to that of transesterified DPNR (TE-DPNR). The addition of diammonium hydrogen phosphate into DPNR-latex in order to remove Mg2+ ions yielded a slight decrease in molecular weight and a slight shift in MWD. The phospholipids are expected to link with mono- and diphosphate groups at the alpha-terminal by hydrogen bonding and/or ionic linkages. The decrease in the molecular weight and Huggins' k' constant of DPNR demonstrates the formation of linear molecules after decomposition of branch-points by this treatment, showing that phospholipids participate in the branching formation of NR. The branch-points formed at the alpha-terminus are postulated to originate predominantly by the association of phospholipids via micelle formation of long-chain fatty acid esters and hydrogen bonding between polar headgroups of phospholipids.

Structural characterization of rubber from Hevea brasiliensis (NR) has been carried out to elucidate the biosynthesis mechanism of rubber molecule as well as to find the relationship between physical properties characteristic of NR and its structure. Recent advances of structural studies have provided a series of new information on the structure of long-chain branching based on the result of selective decomposition of branch-points by chemical and enzymatic treatments as well as physical treatment such as polar solvent treatments and washing of NR latex by centrifugation in the presence of a surfactant. The measurement of the resulting rubber with NMR, FTIR, SEC and dilute solution viscosity provided confirming evidence that the initiating terminal (ω-terminal) with an unidentified functional group and phosphate terminal (α-terminal) form branch-points by hydrogen bond, ionic bond or micelle formation of phospholipids linked to both terminal groups. Based on these results, the origin of green properties characteristic of NR has been explained and a new mechanism of storage hardening has been proposed. The purification method by saponification of NR latex developed for the structural characterization has been applied to produce purified NR latex free from Type I allergic reaction. Instantaneous coagulation of saponified latex by the use of a flocculant and formic acid has provided solid saponified NR having good green and cured rubber properties.