JP2017501292A - Preparation of poly α-1,3-glucan ester and film produced therefrom - Google Patents

Abstract

Disclosed herein are poly α-1,3-glucan ester compounds having a degree of substitution of about 0.05 to about 3.0. Also disclosed is a method of producing a poly α-1,3-glucan ester compound and a film produced therefrom.

Description

  This application is a continuation-in-part of US patent application Ser. No. 14 / 136,168, filed Dec. 20, 2013, all incorporated herein by reference in its entirety, each of Dec. 27, 2012. Claims the benefit of US Provisional Patent Application Nos. 61 / 746,328, 61 / 746,335, and 61 / 746,338, both of which have been filed on the same day and are now complete. .

  The present invention is in the field of poly α-1,3-glucan derivatives. Specifically, the present invention relates to poly α-1,3-glucan esters, methods for their preparation, and films produced therefrom.

  In an effort to find new structural polysaccharides using enzymatic synthesis or genetic manipulation of microorganisms or plant hosts, researchers are able to economically manufacture many raw materials based on biodegradable and renewable resources. I have discovered sugars. One such polysaccharide is poly α-1,3-glucan, a glucan polymer characterized by having α-1,3-glycoside linkages. This polymer has been isolated by contacting an aqueous solution of sucrose with a glucosyltransferase enzyme isolated from Streptococcus salivarius (Non-patent Document 1). Films prepared from poly α-1,3-glucan can withstand temperatures up to 150 ° C. and provide advantages over polymers obtained from β-1,4-linked polysaccharides (Non-Patent Document 2).

  Patent Document 1 is a polysaccharide fiber containing hexose units, in which at least 50% of the hexose units in the polymer are S.P. The preparation of polysaccharide fibers linked by α-1,3-glycosidic linkages utilizing the S. salivarius gtfJ enzyme has been disclosed. This enzyme utilizes sucrose as a substrate in a polymerization reaction that produces poly α-1,3-glucan and fructose as final products (Simpson et al., 1995). The disclosed polymers formed liquid crystal solutions when dissolved above the critical concentration in the solvent or mixture containing the solvent. From this solution, continuous, strong, cotton-like fibers were spun and used, which are very suitable for use in textiles.

  Information on the preparation of various derivatives of poly α-1,3-glucan and their use is poor.

  Non-Patent Document 3 discloses the synthesis of poly α-1,3-glucan triacetate using poly α-1,3-glucan extracted from the fruiting body of a fungus, Laetiporus sylphureus. doing. The structure of this polymer was analyzed by X-ray crystallography.

  Non-Patent Document 4 prepared poly α-1,3-glucan triacetate using three different samples of poly α-1,3-glucan. One of the samples was isolated from bacterial extracellular polysaccharide and the other two samples were extracted from fungal fruiting bodies. The structure of these polymers was analyzed by X-ray crystallography.

  The development of new poly alpha-1,3-glucan ester derivatives and methods for preparing such derivatives are desirable given their potential utility in various applications such as film manufacturing.

US Patent 7,000,000 specification

Simpson et al. , Microbiology 141: 1451-1460, 1995. Ogawa et al. , Fiber Differentiation Methods 47: 353-362, 1980. Yui et al. (Int. J. Biol. Macromol. 14: 87-96, 1992) Ogawa et al. (Carb. Poly. 3: 287-297, 1983).

  The present invention is directed to a film comprising a poly alpha-1,3-glucan ester having (a) at least about 0.1 gf / mil tear resistance; or (b) at least one of haze less than about 20%. .

  In another embodiment, the present invention provides: (a) providing a poly α-1,3-glucan ester; (b) contacting the poly α-1,3-glucan ester of (a) with a solvent to obtain a poly making a solution of α-1,3-glucan ester; (c) applying a solution of poly α-1,3-glucan ester to the surface; and (d) evaporating the solvent to produce poly α-1,3 It is directed to a method of preparing a poly α-1,3-glucan ester film comprising providing a 3-glucan ester film.

  The disclosures of all patent and non-patent literature cited herein are hereby incorporated by reference in their entirety.

  As used herein, the term “invention” or “disclosed invention” is not limiting and generally applies to any of the inventions defined in the claims or described herein. These terms are used interchangeably herein.

The terms “poly α-1,3-glucan”, “α-1,3-glucan polymer”, and “glucan polymer” are used interchangeably herein. A poly α-1,3-glucan is a polymer comprising glucose monomer units linked together by glycosidic linkages, wherein at least about 50% of the glycoside linkages are α-1,3-glycoside linkages. Poly α-1,3-glucan is one type of polysaccharide. The structure of poly α-1,3-glucan can be expressed as follows.

  In the present specification, poly α-1,3-glucan that can be used for the preparation of a poly α-1,3-glucan ester compound can be prepared by using a chemical method. Alternatively, it can be prepared by extraction from various organisms such as fungi producing poly α-1,3-glucan. Further alternatives include, for example, U.S. Patent No. 7,000,000 and U.S. Patent Application Publication Nos. 2013/0244288 and 2013/0244287, all of which are incorporated herein by reference. ), Poly α-1,3-glucan can be enzymatically produced from sucrose using one or more glucosyltransferase (gtf) enzymes (eg, gtfJ).

  The terms “glucosyltransferase enzyme”, “gtf enzyme”, “gtf enzyme catalyst”, “gtf”, and “glucan sucrase” are used interchangeably herein. The activity of the gtf enzyme herein catalyzes the reaction of the sucrose substrate to yield the products poly alpha-1,3-glucan and fructose. Other products (by-products) of the gtf reaction include glucose (when glucose is hydrolyzed from the glucosyl-gtf enzyme intermediate complex), various soluble oligosaccharides (DP2-DP7), and leucus (glucosyl- gtf enzyme intermediate complex glucose may be bound to fructose). Leucrose is a disaccharide composed of glucose and fructose linked by α-1,5 linkages. Wild-type glucosyltransferase enzymes generally include a signal peptide, a variable domain, a catalytic domain, and a glucan binding domain (in the N-terminal to C-terminal direction). Gtf herein is classified into glycoside hydrolase family 70 (GH70) according to the CAZy (Carbohydrate-Active EnZymes) database (Cantarel et al., Nucleic Acids Res. 37: D233-238, 2009).

  The percentage of glycosidic linkages that are α-1,3 between the glucose monomer units of the polyα-1,3-glucan used herein to prepare the polyα-1,3-glucan ester compound is , At least about 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% (or any integer value between 50% and 100%) ). Thus, in such embodiments, the poly alpha-1,3-glucan is less than about 50%, 40%, 30%, 20%, 10%, 5%, 4%, 3%, 2%, 1% Or 0% (or any integer value between 0% and 50%) of non-α-1,3 glycosidic linkages.

  The poly α-1,3-glucan used to produce the poly α-1,3-glucan ester compound herein is preferably linear / unbranched. In certain embodiments, the poly alpha-1,3-glucan has no branch points or is less than about 10%, 9%, 8%, 7%, 6% as a percent of glycosidic linkage in the polymer, It has a branching point of 5%, 4%, 3%, 2%, or 1%. Examples of branch points include α-1,6 branch points such as those present in mutan polymers.

  The terms “glycoside linkage” and “glycoside linkage” are used interchangeably herein and refer to a type of covalent bond that attaches a carbohydrate (sugar) molecule to another group, such as another carbohydrate. As used herein, the term “α-1,3-glycoside linkage” refers to the type of covalent bond that connects α-D-glucose molecules to each other via carbons 1 and 3 of adjacent α-D-glucose rings. Point to. This linkage is shown in the poly α-1,3-glucan structure given above. In the present specification, “α-D-glucose” is referred to as “glucose”.

により表すことができる。 The terms “poly α-1,3-glucan ester compound”, “poly α-1,3-glucan ester”, and “poly α-1,3-glucan ester derivative” are used interchangeably herein. . The poly α-1,3-glucan ester compound herein has the structure:

Can be represented by

  In this structural formula, n may be at least 6 and each R may independently be a hydrogen atom (H) or an organic group. The poly α-1,3-glucan ester compounds herein have a degree of substitution of about 0.05 to about 3.0.

  The poly α-1,3-glucan ester compounds disclosed herein are synthetic artificial compounds.

Poly α-1,3-glucan ester compounds are referred to herein as “esters” based on the inclusion of the basic structure —C _G —O—CO—C—, where “—C _G —” It represents carbon 2, 4, or 6 of the glucose monomer unit of the α-1,3-glucan ester compound, and “—CO—C—” is contained in the acyl group.

As used herein, an “acyl group” includes, for example, an acetyl group (—CO—CH ₃ ), a propionyl group (—CO—CH ₂ —CH ₃ ), a butyryl group (—CO—CH ₂ —CH ₂ —CH). ₃ ), pentanoyl group (—CO—CH ₂ —CH ₂ —CH ₂ —CH ₃ ), hexanoyl group (—CO—CH ₂ —CH ₂ —CH ₂ —CH ₂ —CH ₃ ), heptanoyl group (—CO—) CH ₂ —CH ₂ —CH ₂ —CH ₂ —CH ₂ —CH ₃ ) or an octanoyl group (—CO—CH ₂ —CH ₂ —
May be _{CH 2 -CH 2 -CH 2 -CH 2} -CH 3). The carbonyl group (—CO—) of the acyl group is ester-bonded to carbon 2, 4 or 6 of the glucose monomer unit of the poly α-1,3-glucan ester compound.

  With respect to nomenclature, poly α-1,3-glucan ester compounds can be cited herein by referring to the organic acid corresponding to the acyl group in the compound. For example, an ester compound containing an acetyl group can be called poly α-1,3-glucan acetate, an ester compound containing a propionyl group can be called poly α-1,3-glucan propionate, and a butyryl group. An ester compound containing can be referred to as poly α-1,3-glucan butyrate. However, this nomenclature does not refer to the poly α-1,3-glucan ester compound herein as the acid itself.

  As used herein, “poly α-1,3-glucan triacetate” refers to a poly α-1,3-glucan ester compound having a substitution degree with an acetyl group of 2.75 or more.

  The terms “poly α-1,3-glucan monoester” and “monoester” are used interchangeably herein. The poly α-1,3-glucan monoester contains only one type of acyl group. Examples of such monoesters are poly α-1,3-glucan acetate (including acetyl groups) and poly α-1,3-glucan propionate (including propionyl groups).

  The terms “poly α-1,3-glucan mixed ester” and “mixed ester” are used interchangeably herein. The poly α-1,3-glucan mixed ester contains two or more kinds of acyl groups. Examples of such mixed esters include poly α-1,3-glucan acetate propionate (including acetyl and propionyl groups) and poly α-1,3-glucan acetate butyrate (including acetyl and butyryl groups). ).

  The terms “reactant”, “reaction composition”, and “esterification reactant” are used interchangeably herein and include poly α-1,3-glucan, at least one acid catalyst, at least one type of acid catalyst. It refers to a reactant comprising an acid anhydride and at least one organic acid. The reactant is substantially anhydrous. The reactants are placed under conditions (eg, time, temperature) suitable for esterification of one or more hydroxyl groups of the glucose units of the poly α-1,3-glucan with at least acyl groups from acid anhydrides. , Thereby producing a poly α-1,3-glucan ester compound.

  The terms “substantially anhydrous” and “anhydrous” are used interchangeably herein. Substantially anhydrous conditions are those where there is less than about 1.5 wt% or 2.0 wt% water. Such conditions can characterize, for example, reactants or reaction components.

  As used herein, “acid-exchanged” poly α-1,3-glucan has been treated with acid to remove water from the poly α-1,3-glucan. The acid exchange process for producing acid exchanged poly alpha-1,3-glucan includes one or more treatments in which the glucan is placed in an acid (eg, an organic acid) and then removed from the acid. obtain.

As used herein, the term “acid catalyst” refers to any acid that accelerates the progress of the esterification reaction. Examples of acid catalysts are inorganic acids such as sulfuric acid (H ₂ SO ₄ ) and perchloric acid (HClO ₄ ).

The term “anhydride” as used herein refers to an organic compound having two acyl groups bonded to the same oxygen atom. Typically, acid anhydrides herein have the formula (R—CO) ₂ O, where R is a saturated linear carbon chain (up to 7 carbon atoms). Examples of acid anhydrides are acetic anhydride [(CH ₃ —CO) ₂ O], propionic anhydride [(CH ₃ —CH ₂ —CO) ₂ O], and butyric anhydride [
Is a _{(CH 3 -CH 2 -CH 2 -CO} ) 2 O].

The terms “organic acid” and “carboxylic acid” are used interchangeably herein. The organic acid has the formula R—COOH, where R is an organic group and COOH is a carboxyl group. The R groups herein are typically saturated straight chain carbon chains (up to 7 carbon atoms). Examples of the organic acid is acetic acid _(CH 3 -COOH), propionic acid _{(CH 3} -CH 2 -COOH), and butyric acid _{(CH 3 -CH 2 -CH 2 -COOH} ).

  As used herein, the term “degree of substitution” (DoS) refers to the average number of hydroxyl groups substituted in each monomer unit (glucose) of a poly α-1,3-glucan ester compound. Since each monomer unit of poly α-1,3-glucan has three hydroxyl groups, the DoS of the poly α-1,3-glucan ester compound in this specification can be 3 or less.

  “Contacting” herein may be performed by any means known in the art, such as, for example, dissolution, mixing, shaking, or homogenization. When three or more reaction components are in contact with each other, such contact can be performed all at once or stepwise (eg, mixing the two components followed by the third component).

The “molecular weight” of the poly α-1,3-glucan and the poly α-1,3-glucan ester compound in the present specification is expressed as a number average molecular weight (M _n ) or a weight average molecular weight (M _w ). be able to. Alternatively, the molecular weight can also be expressed as daltons, grams / mole, DPw (weight average polymerization degree), or DPn (number average polymerization degree). Various means are known in the art for calculating these molecular weight measurements, such as high pressure liquid chromatography (HPLC), size exclusion chromatography (SEC), or gel permeation chromatography (GPC).

  The terms “percent by volume”, “volume percent”, “vol%”, and “v / v%” are used interchangeably herein. The percent by volume of solute in solution can be determined using the formula: [(volume of solute) / (volume of solution)] × 100%.

  The terms “percent by weight”, “weight percentage (wt%)”, and “weight-weight percentage (% w / w)” are used interchangeably herein. Percent by weight refers to the percentage of mass-based material contained in the composition, mixture, or solution.

  The terms “increased”, “increased”, and “enhanced” are used interchangeably herein. These terms include, for example, at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% over the amount or activity to which the increased amount or activity is compared. %, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, or 200% (or between 1% and 200% Any integer) may refer to a large amount or activity.

により表されるポリα−１，３−グルカンエステル化合物を含む組成物に関する。 Embodiments of the disclosed invention have the structure:

It relates to a composition comprising a poly α-1,3-glucan ester compound represented by the formula:

  With respect to this structural formula, n may be at least 6 and each R may independently be H or an acyl group. Furthermore, the poly α-1,3-glucan ester compound has a degree of substitution of about 0.05 to about 3.0.

  Each R group in the poly α-1,3-glucan ester compound formula may independently be H or an acyl group. The acyl group in the present specification may be, for example, an acetyl group, a propionyl group, a butyryl group, a pentanoyl group, a hexanoyl group, a heptanoyl group, or an octanoyl group. As such, an acyl group may comprise a chain of 2-8 carbons. This chain preferably has no branches at all.

  The poly α-1,3-glucan ester compound in certain embodiments disclosed herein may contain one type of acyl group. For example, one or more R groups ester bonded to a glucose group in the above formula may be a propionyl group. Thus, the R groups in this particular example will be independently hydrogen and propionyl groups. As another example, one or more R groups esterified to a glucose group in the above formula may be an acetyl group. Thus, the R groups in this particular example will be independently hydrogen and acetyl groups. Certain embodiments of the poly α-1,3-glucan ester compounds herein do not have DoS due to acetyl groups of 2.75 or higher.

  Alternatively, the poly α-1,3-glucan ester compounds disclosed herein can contain two or more different types of acyl groups. Examples of such compounds are (i) an acetyl group and a propionyl group (when the poly α-1,3-glucan acetate propionate, R group is independently H, acetyl, or propionyl), or ( ii) It contains two different acyl groups, such as an acetyl group and a butyryl group (when poly α-1,3-glucan acetate butyrate, R group is independently H, acetyl, or butyryl).

  The poly α-1,3-glucan ester compound has a degree of substitution (DoS) of about 0.05 to about 3.0. Alternatively, the DoS of the poly α-1,3-glucan ester compound disclosed herein can be about 0.2 to about 2.0. Further alternatively, the DoS is at least about 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.. 2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, It can be 2.5, 2.6, 2.7, 2.8, 2.9, or 3.0. Those skilled in the art will appreciate that since the poly α-1,3-glucan ester compounds disclosed herein have a degree of substitution of about 0.05 to about 3.0, the R group of the compound cannot be solely hydrogen. Will be understood.

  Instead of quoting DoS values, the wt% of one or more acyl groups in the poly α-1,3-glucan ester compounds herein may be mentioned. For example, the wt% of the acyl group in the poly α-1,3-glucan ester compound is at least about 0.1%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8 %, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41% 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58 %, 59%, or 60%.

  The percentage of glycosidic linkage that is α-1,3 between the glucose monomer units of the poly α-1,3-glucan ester compound is at least about 50%, 60%, 70%, 80%, 90%, 95% 96%, 97%, 98%, 99%, or 100% (or any integer between 50% and 100%). Thus, in such embodiments, the compound is less than about 50%, 40%, 30%, 20%, 10%, 5%, 4%, 3%, 2%, 1%, or 0% (or 0% %, And any integer value between 50% and 50%).

  The skeleton of the poly α-1,3-glucan ester compound disclosed in the present specification is preferably linear / unbranched. In certain embodiments, the compound has no branch points or less than about 10%, 9%, 8%, 7%, 6%, 5%, 4%, as a percent of glycosidic linkage in the polymer, It has a branching point of 3%, 2% or 1%. Examples of branch points include α-1,6 branch points.

  The formula of the poly α-1,3-glucan ester compound in certain embodiments may have an n value of at least 6. Alternatively, n is at least 10, 50, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, 2000, 2100, 2200, 2300, 2400, 2500, 2600, 2700, 2800, 2900, 3000, 3100, 3200, 3300, 3400, 3500, 3600, 3700, 3800, 3900, or 4000 (or any between 10 and 4000) Integer) value.

The molecular weight of the poly α-1,3-glucan ester compound disclosed in the present specification can be measured as a number average molecular weight (M _n ) or a weight average molecular weight (M _w ). Alternatively, molecular weight can be measured as daltons or grams / mole. It may be useful to refer to DP _w (weight average degree of polymerization) or DP _n (number average degree of polymerization) of the poly α-1,3-glucan polymer component of the compound.

The M _n or M _w of the poly α-1,3-glucan ester compounds disclosed herein can be at least about 1000. Alternatively, M _n or M _w can be at least about 1000 to about 600000. Further alternatively, M _n or M _w is, for example, at least about 10,000, 25000, 50000, 75000, 100000, 125000, 150,000, 175000, 200000, 225000, 250,000, 275000, or 300000 (or any between 10,000 and 300,000). Integer).

  The poly alpha-1,3-glucan ester in certain embodiments has a maximum of about 2.00, 2.05, 2.10, 2.15, 2.20, 2.25, 2.30, 2.35, 2.40, 2.45, 2.50, 2.55, 2.60, 2.65, 2.70, 2.75, 2.80, 2.85, 2.90, 2.95, or 3 It may have a DoS with an acetyl group of 0.00. Thus, for example, DoS due to an acetyl group can be up to about 2.00-2.40, 2.00-2.50, or 2.00-2.65. As another example, DoS with an acetyl group is about 0.05 to about 2.60, about 0.05 to about 2.70, about 1.2 to about 2.60, or about 1.2 to about 2. .70. Such poly α-1,3-glucan esters may be monoesters or mixed esters.

  The poly alpha-1,3-glucan ester in certain embodiments is up to about 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40% 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, or 55% of the propionyl group may have wt%. Such poly α-1,3-glucan esters may be monoesters or mixed esters. For mixed esters, poly alpha-1,3-glucan acetate propionate is, for example, up to about 0.1%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8% %, 9%, or 10% acetyl group wt% and propionyl group wt% by any of the propionyl wt% listed above.

  The poly alpha-1,3-glucan ester in certain embodiments is up to about 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18 %, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51% , 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, or 60% butyryl group wt%. In other embodiments, the poly α-1,3-glucan ester has a maximum of about 0.80, 0.85, 0.90, 0.95, 1.00, 1.05, 1.10, 1.15. Or DoS with a butyryl group of 1.20. Such poly α-1,3-glucan esters may be monoesters or mixed esters. For mixed esters, poly alpha-1,3-glucan acetate butyrate is, for example, up to about 0.1%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8% 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25 %, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, or 36% acetyl group wt% and the butyryl wt% listed above May have a wt% of butyryl groups by any of

（式中、
（ｉ）ｎは少なくとも６であり、
（ｉｉ）各Ｒは、独立に、Ｈ又はアシル基であり、且つ
（ｉｉｉ）化合物は、約０．０５〜約３．０の置換度を有する）
により表されるポリα−１，３−グルカンエステル化合物を製造する。この方法により製造されたポリα−１，３−グルカンエステルは、任意選択で単離できる。 The disclosed invention also relates to a method for producing a poly α-1,3-glucan ester compound. The method contacts poly alpha-1,3-glucan with at least one acid catalyst, at least one acid anhydride, and at least one organic acid in a reaction that is substantially anhydrous. Wherein an acyl group derived from an acid anhydride is esterified to a poly α-1,3-glucan, thereby forming the structure:

(Where
(I) n is at least 6,
(Ii) Each R is independently H or an acyl group, and (iii) the compound has a degree of substitution of about 0.05 to about 3.0)
To produce a poly α-1,3-glucan ester compound represented by: The poly α-1,3-glucan ester produced by this method can be optionally isolated.

  The poly α-1,3-glucan is contacted with at least one acid catalyst, at least one acid anhydride, and at least one organic acid in a reactant that is substantially anhydrous. Substantially anhydrous reactants herein contain no water or less than about 0.05, 0.1, 0.2, 0.3, 0.4, 0.5,. 6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9 or 2.0 wt% water. Substantially anhydrous conditions can be obtained by using reaction components that are substantially anhydrous. Reaction components that are not substantially anhydrous can be used in the preparation of the reactants, but can only be used in amounts such that the final reaction formulation is substantially anhydrous.

  Enzymatically prepared formulations of poly alpha-1,3-glucan that can be used in the disclosed esterification reactions typically contain water. The poly α-1,3-glucan is acid exchanged to remove water, thereby making the glucan substantially anhydrous. In certain embodiments, poly alpha-1,3-glucan is acid exchanged with an organic acid (eg, acetic acid, propionic acid, or butyric acid) prior to contacting step (a) to produce poly alpha-1,3. -Water can be removed from the glucan. The acid exchange process herein involves boiling poly α-1,3-glucan in water and removing most of the water by any physical means (eg, filtration, decantation, and / or drying) Can be included in the organic acid and then removed by filtration and / or decantation. Treatment with an organic acid can include stirring the glucan in the acid, but can be performed once, twice, or more times. The amount of organic acid used in each treatment can be, for example, at least about 2-20 times, or 2-10 times the amount of poly alpha-1,3-glucan being treated.

  The poly alpha-1,3-glucan is contacted with at least one acid catalyst in the disclosed reactants. The acid catalyst may be an inorganic acid in certain embodiments. Examples of inorganic acid catalysts that can be included in the reactants herein are sulfuric acid and perchloric acid. Other examples of inorganic acid catalysts include hydrochloric acid, phosphoric acid, nitric acid, boric acid, hydrofluoric acid, hydrobromic acid, sulfonic acid, any mineral acid, and any combination thereof. The acid catalysts herein are typically commercially available in concentrated (eg, purity> 95%, 96%, 97%, 98%, or 99%) and / or substantially anhydrous forms. Can be obtained. For example, sulfuric acid for use in the reactants herein can be at least about 95-98% pure. Alternatively, the acid catalyst can be provided dissolved in an organic acid such as acetic acid. An example of such a solution is perchloric acid (0.1N) in acetic acid. The amount of acid catalyst in the reactant is, for example, at least about 0.005, 0.0075, 0.01, 0.025, 0.05, 0.075, 0.1, 0.25, 0.5, It can be 0.75, 1.0, 1.25, 1.5, 1.75, or 2.0 wt%.

  The poly α-1,3-glucan contacts at least one acid anhydride in the disclosed reaction. Examples of acid anhydrides that can be included in the reactants herein include acetic anhydride, propionic anhydride, butyric anhydride, pentanoic anhydride, hexanoic anhydride, heptanoic anhydride, octanoic anhydride, And phthalic anhydride. Any combination of these can also be used in the reactants herein (eg, acetic anhydride and propionic anhydride, acetic anhydride and butyric anhydride, propionic anhydride and butyric anhydride). The acid anhydrides herein are typically commercially available in concentrated form (eg,> 95%, 96%, 97%, 98%, or 99% purity) and / or in substantially anhydrous form. Can be obtained. For example, acetic anhydride, propionic anhydride, and / or butyric anhydride for use in the reactions herein can be at least about 97%, 98%, or 99% pure. The amount of acid anhydride in the reactant is, for example, at least about 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, or 70 wt% (or 10 wt% and 70 wt%). Any integer value between). In certain embodiments, the amount of acetic anhydride in the reaction can be at least about 20-45 wt%. In other embodiments, the amount of propionic anhydride or butyric anhydride can be at least about 40-50 wt%.

  The poly α-1,3-glucan contacts with at least one organic acid in the disclosed reactants. Examples of organic acids that can be included in the reactants herein include acetic acid, propionic acid, butyric acid, pentanoic acid, hexanoic acid, heptanoic acid, octanoic acid, and phthalic acid. The amount of organic acid in the reactant is, for example, at least about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, or 80 wt% (or 5 wt %, And any integer value between 80 wt%).

  Typically, the one or more acid anhydrides used in the reactants herein are selected based on the type of esterification desired. For example, if esterification of a poly α-1,3-glucan with an acetyl group, propionyl group, and / or butyryl group is desired, acetic anhydride, propionic anhydride, and / or butyric anhydride respectively. Are included in the reactants. The selected acid anhydride is the main acyl group source in the disclosed esterification process. Nevertheless, the acyl group for esterification can also be derived from one or more organic acids contained in the reactants. For example, an acetyl group, a propionyl group, a butyryl group, a pentanoyl group, a hexanoyl group, a heptanoyl group, and an octanoyl group can be derived from acetic acid, propionic acid, butyric acid, pentanoic acid, hexanoic acid, heptanoic acid, and octanoic acid, respectively. A reactant containing a particular acid anhydride typically also includes an organic acid corresponding to the acid anhydride.

  In certain embodiments of the disclosed reactants, the acid anhydride is one or more of acetic anhydride, propionic anhydride, or butyric anhydride; the organic acid is one of acetic acid, propionic acid, or butyric acid That's it. For example, (i) poly α-1,3-glucan acetate can be prepared using acetic anhydride and acetic acid; (ii) poly α-1,3-glucan propio using propionic anhydride and propionic acid. (Iii) can prepare poly alpha-1,3-glucan butyrate using butyric anhydride and butyric acid; (iv) propionic anhydride, propionic acid, acetic anhydride and optionally (V) Propionic acid anhydride, propionic acid, and acetic acid can be used to prepare poly α-1,3-glucan acetate propionate using acetic acid at Pionates can be prepared; (vi) Poly α-1,3-glucan acetate butyrate can be prepared using butyric anhydride, butyric acid, acetic anhydride, and optionally acetic acid; (vii) An acid anhydride, and butyric acid, a poly-alpha-1,3-glucan acetate butyrate using acetic acid can be prepared. In a reactant comprising acetic acid with propionic acid or butyric acid, the amount of acetic acid can be, for example, about 5-10, 5-20, or 5-30 wt%.

  The reactants for making mixed esters (eg, poly α-1,3-glucan acetate propionate, poly α-1,3-glucan acetate butyrate) are typically acyls where higher DoS is desirable. More acid anhydrides with groups and lower DoS, where less acid anhydrides and / or corresponding organic acids are desirable. For example, to produce poly α-1,3-glucan acetate propionate having a higher DoS of propionyl group compared to acetyl group, more propionic anhydride compared to the amount of acetic anhydride and / or acetic acid Are included in the reactants. The DoS in the mixed ester can also be controlled by the order in which the acid anhydride is added to the reaction already containing the acid catalyst. For example, in preparing a formulation for producing poly alpha-1,3-glucan acetate propionate, propionic anhydride is added prior to acetic anhydride (to a formulation that already contains an acid catalyst). If so, a higher DoS of the propionyl group can be expected.

  The acid anhydride selected for the reactants herein may correspond to the organic acid used to prepare the acid exchanged poly alpha-1,3-glucan. For example, if the reactant comprises propionic anhydride, the acid exchange process can be carried out with propionic acid. Alternatively, the acid anhydride selected for the reactants herein may be different from the organic acid used in the preparation of the acid exchanged poly α-1,3-glucan. For example, if the reactant includes propionic anhydride, the acid exchange process can be performed with acetic acid.

  The reactants herein can include components in addition to poly α-1,3-glucan, acid catalyst, acid anhydride, and organic acid. For example, one or more organic solvents such as methylene chloride can be included in the reaction. An organic solvent such as methylene chloride can be included in the reaction (eg, producing glucan triacetate), for example, at about 30-40 wt%.

  The components of the reactants herein can be added in any order. For example, poly alpha-1,3-glucan, acid catalyst, and organic acid can be mixed first, and then acid anhydride can be added to the mixture. As another example, an acid anhydride and an organic acid can be mixed first, followed by the addition of poly α-1,3-glucan and an acid catalyst to the mixture. As yet another example, the acid catalyst and organic acid can be mixed first, after which poly alpha-1,3-glucan and acid anhydride can be added to the mixture. In certain embodiments, poly alpha-1,3-glucan and another component (eg, acid catalyst or acid anhydride) are added in a sequential order to the mixture containing the other reaction components.

  Cooling can be applied during the various stages of preparing the reactants herein. The terms “cool” and “chill” are used interchangeably herein and refer to reducing the temperature of a reactant or mixture to a lower temperature. Cooling can be performed by any means known in the art, such as an ice bath or an industrial cooling system. Step (a) of preparing the reactants may be carried out after the preparation of the reactants (ie, including all of the poly alpha-1,3-glucan, acid catalyst, acid anhydride, and organic acid) at about 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, or 20 ° C, or cooling to about 12-18 ° C. Alternatively, in step (a), the mixture comprising poly α-1,3-glucan, acid catalyst, and organic acid is cooled (eg, to any of the above temperatures), and then the acid anhydride is converted to the cooled mixture. Adding may be included. Further alternatively, in step (a), the mixture comprising the acid anhydride and the organic acid is cooled (eg, to any of the above temperatures), and then the poly α-1,3-glucan and the acid catalyst are converted to the cooled mixture. Adding may be included. Further alternatively, in step (a), the mixture comprising the acid catalyst and the organic acid is cooled (eg, to any of the above temperatures), and then the poly α-1,3-glucan and the acid anhydride are converted to the cooled mixture. Adding may be included. The reactants can optionally be held for about 1 to 10 minutes at any of the lower temperature points described above after their initial preparation.

  The reactants can then be (i) left under ambient temperature conditions without direct heat and / or (ii) any means known in the art (eg, water bath, industrial heater, Or it can heat directly using an electric heater. Ambient temperature conditions can be maintained, for example, up to about 30, 60, 120, 240, 360, or 480 minutes (or any integer value between 30 and 480 minutes). Alternatively, ambient temperature conditions can be maintained for a maximum of about 24, 48, or 72 hours. As used herein, the term “ambient temperature” refers to a temperature of about 15-30 ° C. or 20-25 ° C. (or any integer between 15 ° C. and 30 ° C.). Reactant heating can be, for example, up to about 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, or 80 ° C. (or any integer value between 30 ° C. and 80 ° C.), about It can be 30-60 ° C, or about 30-50 ° C. Such heating can be performed in stages if desired. For example, the reaction can be first heated to about 35 ° C and then to about 39-50 ° C. Applying the highest reaction temperature (eg, about 36-43 ° C.), poly α-1,3-glucan propionate, poly α-1,3-glucan acetate propionate, or poly α-1,3- In certain embodiments, such as when producing glucan acetate butyrate, an excessive decrease in the molecular weight of the poly α-1,3-glucan ester can be prevented. The temperature after heating to any of the above temperatures can be maintained, for example, for about 20-30, 20-40, 20-60 minutes, or up to about 40, 60, 80, 100, 120, or 140 minutes. If heating is performed in stages, the initial temperature point can be maintained, for example, for about 20-40 minutes. In embodiments where the reactants are subjected to ambient temperature conditions without direct heat, the reactants can be subsequently heated to any of the above temperatures and durations, if desired. The reactants typically do not contain any solid material after any of the above temperature treatments (ambient temperature and / or heating), but may be viscous.

  The reactants can optionally be cooled after any of the above temperature treatments (ambient temperature and / or heating). For example, the reactants can be cooled to about 18, 19, 20, 21, 22, 23, 24, or 25 ° C, about 20-30 ° C, or about 20-40 ° C. The reactants heated to 60-80 ° C can typically be cooled to about 35-45 ° C. The temperature of the reactants when cooled can be maintained, for example, for about 5 to 10 minutes.

  Optionally, the reactants herein can be maintained under an inert gas (eg, nitrogen). As used herein, the term “inert gas” refers to a gas that does not undergo a chemical reaction under a set of given conditions, such as those disclosed for preparing the reactants herein.

  The reactants can optionally be quenched after any of the above temperature treatments (ambient temperature and / or heating) and cooling treatment. Quenching of the reactant can be accomplished by adding an acid, base, or specific salt to the reaction. Various acids, bases, and salts useful for quenching the reactants include acetic acid (eg, about 50-70 wt%), any other mineral or organic acid (eg, about 50-70 wt%), Examples include, but are not limited to, magnesium acetate (eg, about 20-25 wt%), sodium hydroxide, potassium hydroxide, calcium hydroxide, sodium bicarbonate, sodium carbonate, and combinations thereof. In certain embodiments of producing poly alpha-1,3-glucan acetate, the reactants are quenched with acetic acid (eg, about 50 or 70 wt%) or magnesium acetate (eg, about 20-25 wt%).

  The quenched reaction can optionally be heated to about 40 ° C. to 150 ° C. for up to 48 hours. For example, in a process for producing poly alpha-1,3-glucan acetate, the quenched reaction can be heated to about 100 ° C. for up to about 20-40 minutes (eg, 25-30 minutes). Optionally, water may be added to the reaction (quenched or unquenched), then it is about 40 ° C. to 150 ° C. (eg, about 100 ° C.) for up to about 20-40 minutes. (E.g., 25-30 minutes) and heated to reduce the DoS of the acyl group by hydrolysis. Such a heat / water treatment step may be useful to reduce DoS in the process of producing poly alpha-1,3-glucan acetate.

  The poly α-1,3-glucan ester compound produced by the reaction herein can be precipitated using an agent that is a non-solvent for the poly α-1,3-glucan ester compound. For example, deionized water and / or methanol can be added to the reaction solution in an amount sufficient to precipitate the poly α-1,3-glucan ester compound. The precipitation herein may further comprise mixing the reaction solution and the non-solvent by any means known in the art, such as an air driven blender.

  The precipitated poly α-1,3-glucan ester compound is washed twice or more with water, followed by washing in a bicarbonate (eg, sodium bicarbonate) solution (eg, about 5 wt%), Optionally neutralize. The ester compound can then be washed with water once, twice, or more than three times until a neutral pH is obtained. Alternatively, the precipitated poly α-1,3-glucan ester compound is washed with water and a base (for example, diluted alkali hydroxide such as sodium hydroxide, calcium hydroxide, or potassium hydroxide) to adjust the neutral pH. Can be achieved, optionally followed by washing with water. As used herein, the term “neutral pH” refers to a pH that is not substantially acidic or basic (eg, about 6-8, or about 6.0, 6.2, 6.4, 6.6, 6 PH of 7.0, 7.0, 7.2, 7.4, 7.6, 7.8, or 8.0).

  Poly alpha-1,3-glucan esters produced in the disclosed reactants can be isolated. The precipitation process described above can be a step in the isolation process. Isolation utilizes a funnel, centrifuge, press filter, or any other method or apparatus known in the art that can remove liquid from a solid before or after the neutralization and / or washing step. And can be carried out with the precipitated product. The isolated poly alpha-1,3-glucan ester product can be utilized using any method known in the art, such as vacuum drying, air drying (eg, about 16-35 ° C.), or lyophilization. Can be dried.

  Any of the above esterification reactions can be repeated utilizing the poly α-1,3-glucan ester product as a starting material for further modifications. This approach may be suitable for increasing the DoS of the acyl group and / or for adding one or more different acyl groups to the ester product.

  The structure, molecular weight, and DoS of the poly alpha-1,3-glucan ester product can be determined using various physiochemical analyzes known in the art, such as NMR spectroscopy and size exclusion chromatography (SEC). I can confirm.

  The percentage of glycosidic linkages that are α-1,3 between the glucose monomer units of the polyα-1,3-glucan used to prepare the poly α-1,3-glucan ester compounds herein are: , At least about 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% (or any integer value between 50% and 100%) ). Thus, in such embodiments, the poly alpha-1,3-glucan is less than about 50%, 40%, 30%, 20%, 10%, 5%, 4%, 3%, 2%, 1% Or 0% (or any integer value between 0% and 50%) of non-α-1,3 glycosidic linkages.

  The poly α-1,3-glucan used to prepare the poly α-1,3-glucan ester compound herein is preferably linear / unbranched. In certain embodiments, the poly α-1,3-glucan has no branch points or less than about 10%, 9%, 8%, 7%, 6% as a percent of glycosidic linkage in the polymer, It has a branching point of 5%, 4%, 3%, 2%, or 1%. Examples of branch points include α-1,6 branch points.

The M _n or M _w of the poly α-1,3-glucan used to prepare the poly α-1,3-glucan ester compounds herein can be at least about 500 to about 300,000. Further alternatively, M _n or M _w is, for example, at least about 10,000, 25000, 50000, 75000, 100000, 125000, 150,000, 175000, 200000, 225000, 250,000, 275000, or 300000 (or any between 10,000 and 300,000). Integer).

Disclosed herein is a process for producing poly α-1,3-glucan acetate having a DoS of 0.05-2.70 using poly α-1,3-glucan triacetate. The triacetate used in this process can be produced, for example, by any of the processes described above. This process involves contacting poly α-1,3-glucan triacetate with acetic acid and water to form a formulation, and applying a vapor pressure of about 3-10 kg / cm ² to the formulation to bring the temperature to about Including raising to 260 ° C. This process produces poly alpha-1,3-glucan acetate with a DoS of 0.05-2.70. Such a decrease in DoS results from the hydrolysis of a portion of the acetyl group of poly α-1,3-glucan triacetate. The poly α-1,3-glucan acetate produced by this method can be optionally isolated.

  The poly α-1,3-glucan triacetate can optionally be washed and / or has a neutral pH prior to use in this process. Poly alpha-1,3-glucan triacetate can be contacted with acetic acid and water by first dissolving glucan triacetate in acetic acid and then adding water to the solution. In certain embodiments, the amount of acetic acid in the formulation can be about 75, 80, 85, or 90 wt%, and the amount of water in the formulation can be up to about 1, 2, 3, 4, It may be 5, 6, 7, 8, 9, or 10 wt%.

The formulation comprising poly α-1,3-glucan triacetate, acetic acid, and water is then exposed to a vapor pressure of about 3-10 kg / cm ² to raise its temperature to about 260 ° C. This step can optionally be performed in a pressure vessel, such as a perreactor, autoclave, or any other pressure vessel known in the art. For example, a vapor pressure of about 4, 5, or 6 kg / cm ² can be utilized to increase the temperature of the formulation to about 140-160 ° C. (eg, 150 ° C.). This elevated temperature can be maintained for about 30, 40, 50, 60, or 70 minutes, after which the applied pressure can be further increased to about 7, 8, or 9 kg / cm ² . After reaching this increased pressure, the temperature can be cooled to ambient temperature.

  Poly α-1,3-glucan acetate having a DoS of 0.05-2.70 is pressure treated / heat treated using any of the previously disclosed precipitation, washing, and isolation steps. Can be isolated from

  Various types of films can be prepared using poly alpha-1,3-glucan esters formed utilizing the various methods described above. Poly α-1,3-glucan esters prepared according to the disclosed method can be dissolved in one or more solvents to give a solution of poly α-1,3-glucan esters. As used herein, the term “solution of poly α-1,3-glucan ester” refers to a poly α-1,3-glucan ester dissolved in one or more solvents. Useful solvents for this purpose include methylene chloride (dichloromethane), methanol, chloroform, tetrachloroethane, formic acid, acetic acid, nitrobenzene, bromoform, pyridine, dioxane, ethanol, acetone, alcohols, monochlorobenzene, benzene, and toluene. There are, but are not limited to, aromatic compounds, esters such as ethyl acetate and propyl acetate, ethers such as tetrahydrofuran, methyl cellosolve, and ethylene glycol monomethyl ether, or combinations thereof. In one embodiment, poly α-1,3-glucan acetate is dissolved in acetone to prepare a solution of poly α-1,3-glucan acetate. This solution is then applied to the surface and the solvent is evaporated to form a film of the desired thickness. A suitable surface for this application may be, but is not limited to, glass, Teflon®, plastic, or various types of substrates. Methods for making films from the solutions described above are well known in the art and include, but are not limited to, solution pouring, spin coating, thermal spraying, and conventional spraying. In one embodiment, a solution of poly alpha-1,3-glucan ester is fluted on a glass plate.

The tear resistance, tensile strength, and temperature stability of the poly α-1,3-glucan ester film can be measured by methods well known in the art. As used herein, the term “tear resistance” is defined as a measure of how much a film can withstand the effect of tearing. As used herein, the term “tensile strength” refers to the maximum tension a material can withstand without tearing. A suitable tear resistance of the poly α-1,3-glucan ester film disclosed herein may be at least 0.1 gf / mil. The tensile strength of a film suitable for the disclosed invention can be at least 4 kgf / mm ² . In one embodiment, the poly α-1,3-glucan diacetate film has a tear resistance of 2 to 2.4 gf / mil and a tensile strength of 3.97 to 4.8 kgf / mm ² . In another embodiment, the tear resistance is from 1.4 to 3.1 gf / mil and the tensile strength is from 4.98 to 6.44 kgf / mm ² .

  The haze and transmittance of the poly α-1,3-glucan ester film can be determined by methods well known in the art. As used herein, the term “cloudiness” refers to the percentage of light that deviates more than 2.5 degrees from the direction of incident light. A lower haze value corresponds to better transparency. As used herein, the term “transmittance” refers to the rate of incident light of a specified wavelength that passes through a film. Films suitable for poly α-1,3-glucan acetate films in this application can have a haze of up to 20% and a transmittance of at least 80%. In one embodiment, the haze is 6.2% and the transmittance is 94.6%.

  The rate at which the film is produced is determined by adding a weak solvent, such as methanol and cyclohexane, ethanol and n-butanol, or a large amount of methanol or ethanol to methylene chloride and adding to the poly α-1,3-glucan ester solution. It can be increased by accelerating the solidification rate. By controlling the surface temperature and shrinkage percentage of the film, the degree of plane orientation and crystallinity can be controlled.

  The disclosed invention is further defined in the following examples. It should be understood that these examples illustrate certain preferred embodiments of the present invention, but are provided for illustration only. From the above discussion and these examples, one of ordinary skill in the art can ascertain the essential features of the present invention, and various modifications and improvements of the present invention can be made without departing from the spirit and scope thereof. It can be adapted to various applications and conditions.

Abbreviations “mL” is milliliters; “g” is grams; “DI water” is deionized water; “μL” is microliters; “° C.” is degrees Celsius; “mg” is “TFA” is trifluoroacetic acid; “Hz” is hertz; “MHz” is megahertz; “ppm” is parts per million; “HFIP” is hexafluoro-2- “TFA-d” is deuterated trifluoroacetic acid and “kgf” is kilogram weight.

Materials Sulfuric acid, acetic acid, and sodium bicarbonate were obtained from EMD Chemicals (Billerica, MA). Acetic anhydride was obtained from Acros Organics (Pittsburgh, PA). 0.1N perchloric acid in butyric acid, butyric anhydride, propionic anhydride, and acetic acid was obtained from Sigma Aldrich (St. Louis, MO). Propionic acid was obtained from JT Baker (Center Valley, PA). Magnesium acetate was obtained from Alfa Aesar (Ward Hill, Mass.). Unless otherwise noted, all acids and anhydrides used herein were free of water or substantially free of water.

Preparation of poly alpha-1,3-glucan Poly alpha-1,3-glucan is a gtfJ enzyme formulation as described in US 2013/0244288, which is incorporated herein by reference in its entirety. Was used to prepare.

¹ H Nuclear Magnetic Resonance (NMR) Method for Determining the Degree of Substitution of Poly α-1,3-glucan Acetate Derivative The degree of substitution (DoS) of the poly α-1,3-glucan acetate ester derivative is determined using ¹ H NMR. Determined using. Approximately 20 mg of the derivative sample was weighed into a vial on an analytical balance. The vial was removed from the balance and 0.7 mL of TFA-d was added to the vial. A magnetic stir bar was added to the vial and the mixture was stirred until the solid sample was dissolved. Then 0.3 mL of deuterated benzene (C ₆ D ₆ ) was added to the vial to give a better NMR lock signal than TFA-d would give. Using a glass pipette, a 0.8 mL portion of the solution was transferred to a 5 mm NMR tube. Quantitative ¹ H NMR spectra were acquired using an Agilent VNMRS 400 MHz NMR spectrometer equipped with a 5 mm Autoswitchable Quad probe. Spectra were acquired using a spectral window of 399.945 MHz, a spectral window of 6410.3 Hz, an acquisition time of 1.278 seconds, and a 10 second delay between pulses and 124 pulses. Time domain data was transformed using 0.78 Hz exponential multiplication.

  Two regions of the resulting spectrum were integrated: 3.1 ppm to 6.0 ppm giving an integration of seven poly α-1,3-glucan protons and 1.4 ppm to 2.7 ppm giving an integration of three acetyl protons. . The degree of acetylation was calculated by dividing one third of the acetyl proton integration area by one seventh of the poly α-1,3-glucan proton integration area.

¹ H NMR Method for Determining the Degree of Substitution of Poly α-1,3-glucan Propionate Derivatives DoS of poly α-1,3-glucan propionate ester derivatives was determined using ¹ H NMR. . Approximately 20 mg of the derivative sample was weighed into a vial on an analytical balance. The vial was removed from the balance and 0.7 mL of TFA-d was added to the vial. A magnetic stir bar was added to the vial and the mixture was stirred until the solid sample dissolved. Then 0.3 mL of deuterated benzene (C ₆ D ₆ ) was added to the vial to give a better NMR lock signal than TFA-d would give. Using a glass pipette, a 0.8 mL portion of the solution was transferred to a 5 mm NMR tube. Quantitative ¹ H NMR spectra were acquired using an Agilent VNMRS 400 MHz NMR spectrometer equipped with a 5 mm Autoswitchable Quad probe. Spectra were acquired using a spectral window of 399.945 MHz, a spectral window of 6410.3 Hz, an acquisition time of 1.278 seconds, and a 10 second delay between pulses and 32 pulses. Time domain data was transformed using 1.0 Hz exponential line broadening and the benzene solvent peak was set to 7.15 ppm.

  For poly α-1,3-glucan propionate samples, three regions of the resulting spectrum: 3.3 ppm to 6.0 ppm giving integration of seven poly α-1,3-glucan protons; methylene of propionyl group 1.9 ppm to 2.7 ppm giving the integration of the methyl group of the group plus acetyl group; and 0.8 ppm to 1.3 ppm giving the integration of the methyl group of the propionyl group.

  The DoS due to the propionyl group was calculated by dividing the integral value of the methyl group of the propionyl group by 3. Next, the integral value of the methylene group of the propionyl group was calculated by multiplying the integral value of the methyl group of the propionyl group by 0.666. This value was then subtracted from the integral of the methylene group of the propionyl group plus the methyl group of the acetyl group to give the integrated value of the methyl group of the acetyl group.

¹ H NMR method for determining the degree of substitution of poly α-1,3-glucan mixed ester derivatives The DoS of poly α-1,3-glucan mixed ester derivatives was determined using ¹ H NMR. Approximately 20 mg of the derivative sample was weighed into a vial on an analytical balance. The vial was removed from the balance and 0.7 mL of TFA-d was added to the vial. A magnetic stir bar was added to the vial and the mixture was stirred until the solid sample dissolved. Then 0.3 mL of deuterated benzene (C ₆ D ₆ ) was added to the vial to give a better NMR lock signal than TFA-d would give. Using a glass pipette, a 0.8 mL portion of the solution was transferred to a 5 mm NMR tube. Quantitative ¹ H NMR spectra were acquired using an Agilent VNMRS 400 MHz NMR spectrometer equipped with a 5 mm Autoswitchable Quad probe. Spectra were acquired using a spectral window of 399.945 MHz, a spectral window of 6410.3 Hz, an acquisition time of 1.278 seconds, and a delay between pulses of 10 seconds and 32 pulses. Time domain data was converted using exponential line broadening at 1.0 Hz and the benzene solvent peak was set at 7.15 ppm.

  In the poly α-1,3-glucan acetate propionate sample, three regions of the resulting spectrum: 3.3 ppm to 6.0 ppm giving integration of seven poly α-1,3-glucan protons; of the propionyl group 1.9 ppm to 2.7 ppm giving the integration of the methyl group of the methylene group plus acetyl group; and 0.8 ppm to 1.3 ppm giving the integration of the methyl group of the propionyl group.

  The DoS due to the propionyl group on the glucan was calculated by dividing the integral of the methyl group of the propionyl group by 3. Next, the integral value of the methylene group of the propionyl group was calculated by multiplying the integral value of the methyl group of the propionyl group by 0.666. This value was then subtracted from the integral of the methylene group of the propionyl group plus the methyl group region of the acetyl group to give the integrated value of the methyl group of the acetyl group. Finally, the acetyl group integral value was divided by 3 to obtain the degree of acetylation.

  For poly α-1,3-glucan acetate butyrate samples, three regions of the resulting spectrum: 3.3 ppm to 6.0 ppm giving integration of seven poly α-1,3-glucan protons; carbonyl of the butyryl group 1.9 ppm to 2.6 ppm giving the integration of the methyl group of the methylene group plus acetyl group in the α of the group; and 0.6 ppm to 1.0 ppm giving the integration of the methyl group of the butyryl group.

  The DoS due to the butyryl group on the glucan was calculated by dividing the integral of the methyl group of the butyryl group by 3. Next, the integral value of the methylene group of the butyryl group was calculated by multiplying the integral value of the methyl group of the butyryl group by 0.666. This value was then subtracted from the integration of the butyryl methylene group plus the methyl group of the acetyl group to give the integrated value of the methyl group of the acetyl group. Finally, the acetyl group integral value was divided by 3 to obtain the degree of acetylation.

Determination of degree of polymerization The degree of polymerization (DP) was determined by size exclusion chromatography (SEC). The poly α-1,3-glucan ester was dissolved in HFIP (2 mg / mL) while shaking at 45 ° C. for 4 hours. The chromatographic system utilized was three online detectors: a Waters differential refractometer 2410, a Wyatt Technologies (Santa Barbara, Calif.) Multi-angle light scattering photometer Heleos ™ 8+, and a Wyatt Technologies differential capillary viscometer ( differential Capillary Viscometer) Alliance ™ 2695 separation module of Waters Corporation (Milford, Mass.) connected to ViscoStar ™. The columns used for SEC were two Shodex (Showa Denka America, New York) GPC HFIP-806M ™ styrene-divinylbenzene columns and one Shodex GPC HFIP-804M ™ styrene-divinylbenzene column. The mobile phase was redistilled HFIP containing 0.01 M sodium trifluoroacetate. The chromatogram conditions utilized were 50 ° C. in the column and detector compartments, 40 ° C. in the sample and injector compartments, a flow rate of 0.5 mL / min, and an injection volume of 100 μL. The software package used for data reduction was Wyatt's Astra version 6 (triple detection method with column calibration).

Example 1
Preparation of acid-exchanged poly α-1,3-glucan This example shows the preparation of acid-exchanged poly α-1,3-glucan that can be used for the preparation of ester derivatives of polyα-1,3-glucan. Will be explained.

  Acid-exchanged poly α-1,3-glucan was prepared by placing 10 g of poly α-1,3-glucan in a 250 mL glass beaker with 150 mL of DI water. This mixture was boiled on a hot plate for 1 hour, after which the poly α-1,3-glucan was recovered by vacuum filtration. The poly α-1,3-glucan is then subjected to two acid exchange steps which are stirred with 100 mL of glacial acetic acid at room temperature followed by vacuum filtration to give acid exchanged poly α-1,3-glucan. Glucan was produced.

  Other forms of acid exchanged poly alpha-1,3-glucan were also prepared according to the above process using propionic acid or butyric acid instead of acetic acid.

  Various poly α-1,3-glucan ester derivatives were prepared using acid-exchanged poly α-1,3-glucan prepared by these techniques in some of the following examples. Since the acid exchange process removes water from the poly α-1,3-glucan, even if the acid-exchanged poly α-1,3-glucan is introduced into the esterification reaction product containing the acid anhydride, the acid anhydride Water that can react with is not introduced.

Example 2
Preparation of Poly α-1,3-glucan Acetate This example illustrates the preparation of a glucan ester derivative, poly α-1,3-glucan acetate.

Acid exchanged poly alpha-1,3-glucan (10 g) prepared in Example 1 using acetic acid was added to 180 mL of a 500 mL round bottom flask equipped with a magnetic stir bar, thermocouple, and condenser. To a mixture containing acetic acid and 1.84 g sulfuric acid. This mixture was stirred at ambient temperature for 1 minute, after which acetic anhydride (50 mL) was added to the mixture. The reaction was allowed to proceed for 30 minutes at ambient temperature, then heated in a 35 ° C. water bath for 20 minutes, followed by heating to 50 ° C. for 30 minutes. The resulting reaction formulation did not contain any solids. The reaction was then removed from the water bath and cooled to 42 ° C. for 15 minutes. The reaction was then quenched with 25 mL of 70% acetic acid and stirred for 40 minutes. Poly α-1,3-glucan acetate was precipitated using an air driven blender and DI water. The solid was washed twice with water for 30 minutes followed by one wash with 5% sodium bicarbonate. The poly α-1,3-glucan acetate solid was then finally washed with water (2 water washes) until neutral pH was achieved. The solid was collected by vacuum filtration, dried under vacuum and characterized by NMR and SEC. This method yielded poly alpha-1,3-glucan acetate with a DoS of 2.3 and _Mn of 29170.

  In this way, an ester derivative, poly α-1,3-glucan acetate, was prepared and isolated.

Example 3
Additional Preparation of Poly α-1,3-glucan Acetate This example illustrates the production of a glucan ester derivative, poly α-1,3-glucan acetate, utilizing a variety of reaction conditions.

Acid exchanged poly alpha-1,3-glucan was prepared as in Example 1 using acetic acid. A mixture of 180 mL acetic acid and 0.08 g concentrated sulfuric acid was prepared in a 500 mL round bottom flask equipped with a magnetic stir bar and thermocouple. The mixture was cooled to 18 ° C. Acid exchanged poly α-1,3-glucan (10 g) was slowly added to the cooled mixture and stirred for 1 minute. Acetic anhydride (50 mL) was then added to the mixture. The reaction was allowed to proceed for 10 minutes without heating and then heated in a 35 ° C. water bath for 20 minutes. The resulting reaction was completely free of solids and was cooled to 22 ° C. over 7 minutes using an ice bath. The reaction was then quenched with 25 mL of 70% acetic acid and stirred for 40 minutes. Poly α-1,3-glucan acetate was precipitated, washed and analyzed as described in Example 2. This process yielded poly alpha-1,3-glucan acetate with a DoS of 2.41 and an _Mn of 73960.

  Different ester products could be formed using different concentrations of reagents. Table 1 below shows different poly α-1,3-glucan acetate esters synthesized using a process similar to the above process but with the specific changes shown in the table. The results in Table 1 show that by changing the reaction conditions and the molecular weight of the poly α-1,3-glucan starting material used in the reaction, the DoS due to the acetyl groups in the ester product and the molecular weight of the product can be changed. Indicates.

  In this way, various forms of ester derivatives, poly α-1,3-glucan acetate, were prepared and isolated.

Example 4
Additional Preparation of Poly α-1,3-glucan Acetate This example describes a process for producing poly α-1,3-glucan acetate having a hydrolysis step.

  Acid exchanged poly alpha-1,3-glucan (28 g) prepared as in Example 1 using acetic acid was added to a mixture containing 93.4 mL acetic acid and 2.24 g concentrated sulfuric acid and mixed. This mixture was added to a 1 L jacketed reaction vessel equipped with an overhead stirrer and thermocouple and cooled to 12 ° C. using a recirculation bath. The reaction mixture was then stirred for 1 minute before acetic anhydride (89 mL) was added. The reaction was heated for 40 minutes using a recirculation bath set at 42 ° C. The reaction at this stage contained no solids and was quenched with 15.25 g (24%) magnesium acetate with excess water to reduce the sulfuric acid content to 2%. The reaction was then heated to 100 ° C. over 25 minutes and then stirred at this temperature for 2 hours. 24% magnesium acetate was added in 5% excess (6.1 g) to completely quench the reaction. Poly α-1,3-glucan acetate was precipitated, washed and analyzed as in Example 2. This process yielded poly alpha-1,3-glucan acetate with a DoS of 2.58.

  Thus, an ester derivative, poly α-1,3-glucan acetate, was prepared using a method incorporating a hydrolysis step.

Example 5
Preparation of poly α-1,3-glucan acetate by hydrolysis of poly α-1,3-glucan triacetate This example illustrates, in part, poly α-1,3 by hydrolysis of poly α-1,3-glucan triacetate. -Describe the preparation of glucan acetate.

  Poly α-1,3-glucan triacetate was first prepared as follows.

Acetic acid (384 mL), acetic anhydride (990 mL), and methylene chloride (890 mL) were mixed. This formulation was added to a 4 L glass reaction vessel equipped with an overhead stirrer and thermocouple and cooled to 12 ° C. Acid exchanged poly alpha-1,3-glucan (130 g) prepared in Example 1 using acetic acid was slowly added to the cooled mixture and stirred for 1 minute. Then, perchloric acid (0.1 N) in acetic acid (180 mL) was added. The reaction was allowed to proceed for 3 hours 35 minutes at ambient temperature. The reaction was free of solids at all and precipitated poly α-1,3-glucan triacetate in addition to an air driven blender containing methanol. The poly α-1,3-glucan triacetate solid thus formed was washed with methanol for 30 minutes, followed by two washes with deionized (DI) water, and once with 5% sodium bicarbonate. Washed. The poly α-1,3-glucan triacetate solid is then finally washed with water until a neutral pH is achieved (2 water washes), collected by vacuum filtration, dried under vacuum, Characterized by NMR and SEC. The produced poly α-1,3-glucan triacetate had a DoS of 3.0 and a M _n of 132300.

The previously prepared poly α-1,3-glucan triacetate was prepared for hydrolysis by first dissolving in 80 mL acetic acid. DI water (4 mL) was then added to the formulation and stirred using a magnetic stir bar until thoroughly mixed. The formulation was then transferred to a perreactor (Parr Instrument Company, Moline, IL). Steam with a pressure of 5 kg / cm ² was blown into the reactor and the temperature was raised to 150 ° C. in 12 minutes. The formulation was kept at this temperature for 50 minutes. Then, the pressure is increased from 5 kg / cm ² to 8.37kg / cm ^2, and then cooling the reaction vessel to ambient temperature. The formulation recovered from the reactor was yellow. However, when DI water was added, a white solid of poly α-1,3-glucan acetate precipitated. The poly α-1,3-glucan acetate was isolated using vacuum filtration, washed and analyzed as in Example 2. This process yielded poly alpha-1,3-glucan acetate with a DoS of 2.4 and _Mn of 44200.

  Thus, poly α-1,3-glucan acetate having a DoS of less than 2.75 was prepared from poly α-1,3-glucan triacetate.

Example 6
Preparation of Poly α-1,3-glucan propionate This example illustrates the preparation of a glucan ester derivative, poly α-1,3-glucan propionate.

The acid exchange poly alpha-1,3-glucan (M _n of _119,130) was prepared as described in Example 1, except that propionic acid was used in place of acetic acid. Propionic acid (8 mL) and sulfuric acid (0.03 g) were mixed in a 250 mL round bottom flask and cooled to 18 ° C. Acid exchanged poly α-1,3-glucan (2 g) was slowly added to the cooled mixture and stirred for 1 minute. Propionic anhydride (10 mL) was then added to the formulation followed by 0.6 mL glacial acetic acid. The reaction was allowed to proceed for 5 minutes without heating, then heated in a 42 ° C. water bath for 1 hour 45 minutes. The maximum temperature was not allowed to exceed 43 ° C. to prevent an excessive decrease in molecular weight. The resulting reaction formulation contained no solids and was cooled to 20 ° C. over 5 minutes using an ice bath. The reaction was then quenched with 4 mL of 50% aqueous acetic acid and stirred for 45 minutes. Poly α-1,3-glucan propionate was precipitated using an air driven blender and DI water. The solid was washed twice with water for 30 minutes followed by once with 5% sodium bicarbonate. The solid of poly α-1,3-glucan propionate was then washed with water until a neutral pH was achieved (2 water washes). The solid was collected by vacuum filtration, dried under vacuum and characterized by NMR and SEC. The solid was identified as poly α-1,3-glucan propionate having 44.1 wt% propionyl group (0 wt% acetyl group) and 59 510 M _n .

  Thus, an ester derivative, poly α-1,3-glucan propionate was prepared and isolated.

Example 7
Preparation of Poly α-1,3-glucan acetate butyrate This example illustrates the preparation of a glucan mixed ester derivative, poly α-1,3-glucan acetate butyrate.

Acid-exchanged poly α-1,3-glucan (10 g) prepared in Example 1 with acetic acid was placed in a 500 mL round bottom flask equipped with a magnetic stir bar, thermocouple, and condenser in 21 mL of glacial acetic acid, To a mixture containing 20 mL butyric acid and 0.09 g sulfuric acid. The mixture was cooled to 18 ° C. using an ice bath and stirred for 1 minute before butyric anhydride (39 mL) was added to the flask. The reaction was allowed to proceed for 10 minutes without heating, then heated in a 35 ° C. water bath for 80 minutes, followed by heating to 39 ° C. for 30 minutes, but the maximum temperature reached was 39 ° C., an excess of product molecular weight. Prevent the decline. The resulting viscous solution contained no solids and the solution was cooled to 20 ° C. for 10 minutes using an ice bath. The reaction was then quenched with 20 mL of 50% aqueous acetic acid and stirred for 40 minutes. Solid poly α-1,3-glucan acetate butyrate was precipitated using an air driven blender and DI water. The solid was washed twice with water for 30 minutes followed by 5% sodium bicarbonate. The solid thus obtained was then finally washed with DI water until a neutral pH was achieved (2 water washes). The solid was collected by vacuum filtration, dried under vacuum and characterized by NMR and SEC. This process yielded a poly α-1,3-glucan acetate butyrate mixed ester having a butyryl DoS of 1.0, an acetyl DoS of 1.3, and a number average molecular weight (M _n ) of 66340.

  Different mixed ester products could be formed using different concentrations of reagents. Table 2 below shows different poly α-1,3-glucan acetate butyrate esters synthesized using a process similar to the above process but with the specific changes shown in the table. The results in Table 2 show that the amount of acetyl and butyryl groups in the mixed ester product and the molecular weight of the product by changing the reaction conditions and the molecular weight of the poly α-1,3-glucan starting material used in the reaction. It can be changed.

  Thus, various forms of mixed ester derivatives, poly α-1,3-glucan acetate butyrate, were prepared and isolated.

Example 8
Preparation of Poly α-1,3-glucan Acetate Propionate This example illustrates the preparation of a glucan mixed ester derivative, poly α-1,3-glucan acetate propionate.

Acid-exchanged poly α-1,3-glucan was prepared as described in Example 1 using acetic acid. A mixture of 35 mL propionic acid and 0.09 g sulfuric acid was prepared in a 500 mL round bottom flask and cooled to 18 ° C. Acid exchanged poly α-1,3-glucan solid (10 g) was slowly added to the cooled mixture and stirred for 1 minute. Propionic anhydride (50 mL) was then added followed by 5 mL of glacial acetic acid. The reaction was allowed to proceed for 10 minutes without heating, then heated in a 30 ° C. water bath for 1 hour followed by heating to 34 ° C. for 10 minutes. The maximum temperature did not exceed 36 ° C. to prevent excessive reduction in product molecular weight. The solution thus obtained contained no solids and the solution was cooled to 20 ° C. for 5 minutes in an ice bath. The reaction was then quenched with 20 mL of 50% aqueous acetic acid and stirred for 40 minutes. Poly α-1,3-glucan acetate propionate was precipitated using an air driven blender and DI water. The solid poly alpha-1,3-glucan acetate propionate product was washed twice with water for 30 minutes followed by one wash with 5% sodium bicarbonate. The solid was then washed with water until a neutral pH was achieved (2 water washes). The solid was collected by vacuum filtration, dried under vacuum and characterized by NMR and SEC. The solid produced was identified as poly α-1,3-glucan acetate propionate containing 17.6 wt% acetyl groups and 32.9 wt% propionyl groups and having a M _n of 64,030.

  Different mixed ester products could be formed using different concentrations of reagents. Table 3 below shows the different poly α-1,3-glucan acetate propionate esters synthesized using a process similar to the above process but with the specific changes shown in the table. The results in Table 3 show the amount of acetyl and propionyl groups in the mixed ester product and the molecular weight of the product by varying the reaction conditions and the molecular weight of the poly α-1,3-glucan starting material used in the reaction. It can be changed.

  Thus, various forms of mixed ester derivatives, poly α-1,3-glucan acetate propionate, were prepared and isolated.

Example 9
Preparation of Poly α-1,3-glucan Triacetate Using Sulfuric Acid as a Catalyst This example illustrates the preparation of poly α-1,3-glucan triacetate using sulfuric acid as a catalyst in the reaction.

Acid exchanged poly alpha-1,3-glucan was prepared as described in Example 1 using acetic acid. A mixture containing 180 mL acetic acid and 1.84 g concentrated sulfuric acid as catalyst was prepared in a 500 mL round bottom flask equipped with an overhead stirrer and thermocouple. Acid exchanged poly α-1,3-glucan (10 g) was slowly added to the mixture and stirred for 1 minute under nitrogen. The mixture was cooled to about 18 ° C. using an ice bath. Acetic anhydride (50 mL) was added to the reaction and then heated to 80 ° C. over 45 minutes and allowed to react at this temperature for 30 minutes. The reaction contained no solids and was cooled to 40 ° C. over 5 minutes using an ice bath. The reaction was then quenched with 25 mL of 70% acetic acid and stirred for 30 minutes. Poly α-1,3-glucan triacetate was precipitated using an air driven blender (Waring, Torrington, CT) and DI water. The solid poly alpha-1,3-glucan triacetate product was washed twice with water for 30 minutes followed by 5% sodium bicarbonate. The solid was then washed with water until a neutral pH was achieved (2 water washes). The solid was collected by vacuum filtration, dried under vacuum and characterized by NMR and SEC. This process yielded 7.8 g of poly α-1,3-glucan triacetate with a DoS of 3.1 and a M _n of 5130. DoS readings above 3.0 probably reflect the integral variation typical of the NMR measurement process.

  Table 4 shows poly α-1,3-glucan triacetate esters of different molecular weights synthesized using a process similar to the above process but with the specific changes shown in the table. The results in Table 4 show that the molecular weight of the product can be changed by changing the reaction conditions and the molecular weight of the poly α-1,3-glucan starting material used in the reaction.

  Thus, various forms of poly α-1,3-glucan triacetate were prepared and isolated in the reaction using sulfuric acid as a catalyst.

Example 10
Preparation of poly α-1,3-glucan triacetate using perchloric acid as catalyst This example illustrates the preparation of poly α-1,3-glucan triacetate using perchloric acid as a catalyst in the reactants. .

Acid exchanged poly alpha-1,3-glucan was prepared as described in Example 1 using acetic acid. A mixture of 384 mL acetic acid, 990 mL acetic anhydride, and 890 mL methylene chloride was prepared in a 4 L glass reaction vessel equipped with an overhead stirrer and thermocouple and cooled to 12 ° C. Acid exchanged poly α-1,3-glucan (130 g) was slowly added to the cooled mixture and stirred for 1 minute. Perchloric acid (0.1N) in acetic acid (180 mL) was then added to the mixture. The reaction was allowed to proceed for 3 hours 35 minutes at ambient temperature. The obtained reaction product did not contain any solids, and the reaction product was added to an air-driven blender containing methanol to precipitate poly α-1,3-glucan triacetate. The solid of poly α-1,3-glucan triacetate was washed with methanol for 30 minutes, followed by washing twice with DI water and once with 5% sodium bicarbonate. The poly α-1,3-glucan triacetate was then washed with water until a neutral pH was achieved (2 water washes). The solid was collected by vacuum filtration, dried under vacuum and characterized by NMR and SEC. This process yielded 221.5 g of poly α-1,3-glucan triacetate with a DoS of 3.2 and a M _n of 132300. DoS readings above 3.0 probably reflect the integral variation typical of the NMR measurement process.

  Table 5 shows the different molecular weight poly α-1,3-glucan triacetate esters synthesized using a process similar to the above process but with the specific changes shown in the table. The results in Table 5 show that the molecular weight of the product can be changed by changing the reaction conditions and the molecular weight of the poly alpha-1,3-glucan starting material used in the reaction.

  Thus, various forms of poly α-1,3-glucan triacetate were prepared and isolated in a reaction using perchloric acid as a catalyst.

Example 11
Preparation of Film Using Poly α-1,3-glucan Acetate Poly α-1,3-glucan acetate prepared as in Example 2 was dissolved in acetone with a 10 wt% mixture to form a solution. The solution was then poured onto a clean glass plate with a film pouring machine and the solvent was evaporated to dryness to give a film. The film was removed from the glass and rinsed with DI water. Table 6 shows the properties of two different poly α-1,3-glucan acetate films prepared using two different samples of poly α-1,3-glucan acetate.

Example 12
Optical Analysis of Poly α-1,3-glucan Acetate Film A sample of the poly α-1,3-glucan acetate film prepared in Example 11 was analyzed for color and haze. The collected spectrum was consistent with ASTM E1164-09a. Spectral bandwidth (SBW) = 1 and wavelength range = 830-360 nm at 1 nm intervals. Table 7 shows the results of this test.

Example 13
Preparation of poly α-1,3-glucan triacetate film Poly α-1,3-glucan triacetate was prepared as described in Example 10. A 10 wt% solution of poly α-1,3-glucan triacetate was prepared by dissolving 10 g in 90 g of methylene chloride: methanol (11.5: 1 v / v). The solution was then fluted onto a clean glass plate with a Gardner Knife (Gardner Lab Inc., Bethesda, MD). The solvent was evaporated to dryness. The film produced after solvent evaporation was removed from the glass and rinsed with DI water. Table 8 summarizes the tensile and tear data for the poly α-1,3-glucan triacetate films produced using this method. It can be seen that variations in the _Mn and DoS of the constituent glucan esters give different properties to the produced films.

Example 14
Thermal Analysis of Poly α-1,3-glucan Triacetate Film The poly α-1,3-glucan triacetate film prepared in Example 13 was analyzed using MDSC and TGA. MDSC measurement uses Q1000TA instrument in N ₂ starting from 0 ° C. with 5-6 mg film, heating rate 3 ° C./min, modulation amplitude 0.48 ° C. and modulation duration 60 seconds. And carried out.

TGA experiments were performed using a Q500TA instrument at ambient temperature to 800 ° C. under N ₂ .

The information given in Table 9 shows the thermal stability / thermal decomposition of the poly α-1,3-glucan triacetate film prepared by the previously disclosed method. Table 9 summarizes the data obtained from MDSC and TGA measurements. It can be seen that variations in the _Mn and DoS of the constituent glucan esters give different properties to the produced films.

Example 15
Optical Analysis of Poly α-1,3-glucan Triacetate Film The poly α-1,3-glucan triacetate film prepared in Example 13 was analyzed for color and haze. Spectra were collected using 1 spectral bandwidth (SBW) and a wavelength range of 830-360 nm at 1 nm intervals to meet ASTM E1164-09a. Table 10 shows the results of optical measurement of the poly α-1,3-glucan triacetate film.

Claims (7)

(A) a tear resistance of at least about 0.1 gf / mil; or (b) a film comprising a poly alpha-1,3-glucan ester having at least one haze of less than about 20%.   The film according to claim 1, wherein the poly α-1,3-glucan ester used is poly α-1,3-glucan acetate. The film of claim 1, having (a) a tear resistance of about 2.0 to about 2.4 gf / mil; or (b) a haze of less than about 10%.   The film according to claim 3, wherein the α-1,3-glucan ester used is poly α-1,3-glucan acetate. (A) providing a poly α-1,3-glucan ester;
(B) contacting the poly α-1,3-glucan ester of (a) with a solvent to make a solution of poly α-1,3-glucan ester;
(C) applying the solution of poly α-1,3-glucan ester to a surface; and (d) evaporating the solvent to give a poly α-1,3-glucan ester film. A method for preparing an α-1,3-glucan ester film.   The process according to claim 5, wherein the poly α-1,3-glucan ester used is poly α-1,3-glucan acetate.   The method of claim 5, wherein the solvent is acetone.