RESSALVA 

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de 22/01/2022. 



          

UNIVERSIDADE ESTADUAL PAULISTA 
“JÚLIO DE MESQUITA FILHO” 

INSTITUTO DE BIOCIÊNCIAS – RIO CLARO unesp 
 

 

 

 

PROGRAMA DE PÓS-GRADUAÇÃO EM CIÊNCIAS BIOLÓGICAS 
(MICROBIOLOGIA APLICADA) 

 

 

 

 

 

CONTRIBUTION OF XYLAN TO PHYSICOCHEMICAL AND BIODEGRADATION 
PROPERTIES IN STARCH-BASED BIOPLASTICS. 

 

 

 

JOÃO VICTOR CARPINELLI MACEDO 
 

 

 

  

 

 

 

 

 

 

 

 

 

Rio Claro 
Julho - 2021 



 

 

PROGRAMA DE PÓS-GRADUAÇÃO EM CIÊNCIAS BIOLÓGICAS 
(MICROBIOLOGIA APLICADA) 

 

 

 

 

 

 

CONTRIBUTION OF XYLAN TO PHYSICOCHEMICAL AND 
BIODEGRADATION PROPERTIES IN STARCH-BASED BIOPLASTICS. 

 

 

 

 

JOÃO VICTOR CARPINELLI MACEDO 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

             
 

 

 

 

 Rio Claro 

Julho - 2021 

Dissertação apresentada ao 
Instituto de Biociências do Câmpus 
de Rio Claro, Universidade 
Estadual Paulista, como parte dos 
requisitos para obtenção do título 
de Mestre em Microbiologia 
Aplicada. 
 
Orientador: Michel Brienzo 



 

 
 

  



 

 

  



 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

I dedicate this work to all my 

family and to everyone who 

helped me get to where I 

am. 



 

 

ACKNOWLEDGEMENT 

 

To my advisor Michel Brienzo, for giving me the opportunity to be part of his students. And 

trusting me to develop an unprecedented project in his laboratory, which together, we got to 

develop, starting a new line of research that will generate many fruits for IPBEN and for 

scientific literature. I acknowledge too all the patience and support offered me throughout the 

master’s, teaching me and helping me to solve the problems related to this project. 

 

To my family, my mother, grandmother, aunts, uncles and cousins for giving me the 

possibility to graduate and get here. In addition to motivating and strengthening me during all 

these years of study. 

 

To all the employees of IPBEN for their services that facilitate our lives in the laboratory and 

colleagues who helped me with so many suggestions, in addition to providing me with good 

times over these two years. 

 

To Marcia Cristina Branciforti from USP São Carlos, her doctoral student Paula Bertolino 

Sanvezzo, Renato Grillo from Unesp in Ilha Solteira and the all the technicians for their help 

in carrying out experiments that we had no access to at IPBEN, in addition to dedicating 

themselves to clarifying us doubts and give suggestions in carrying out this study. 

 

To the postgraduate program in Applied Microbiology, to all its professors and technical 

section for their teaching and support. 

 

To CNPq and Fapesp, which despite suffering a reduction in the budget in recent years, is still 

a competent organization that helps to produce science in the country by investing and 

providing that works like this are developed. Generating more critical individuals, who know 

the relevance of science for the well-being and sustainable development of our country. 

 

The present work was carried out with the support of the National Council for Scientific 

Development and Technological (CNPq) 

 

 

 



 

 

ABSTRACT 

 

Plastic has served society very well for decades, being able to present itself as a light and 

moldable material, mechanically and thermally resistant, inert to chemical products, electrical 

insulator, in addition to having interesting optical characteristics such as its transparency. 

Such characteristics, associated with the low price of its raw material, a residue from the 

processing of petroleum until then little used, made it crucial for the industry and the market, 

which consolidated its presence in society in different countries. However, as it is a resistant 

material, remaining for centuries without fully decomposing, and for having a large 

production in order to meet the needs of a consumer society, it began to accumulate in the 

environment over the years since its creation. Its abundance in the environment causes 

damage to life in different biomes, especially marine environments, in addition to having 

residues that can be toxic and reach humans through the food chain. A possible solution is 

biodegradable materials that use renewable raw materials such as proteins, lipids, and 

polysaccharides, the so-called bioplastics. In this context, the present study aimed to produce 

bioplastics based on starch with different proportions of xylan extracted from sugarcane 

bagasse and glycerol as a plasticizer. For this, their physicochemical properties (solubility, 

hydrophilicity, moisture, water barrier, opacity, crystallinity, and mechanical resistance) were 

evaluated. The biodegradation of the bioplastic was determined by buried and CO2 released. 

As a result, it was observed that the increase in the proportion of xylan made the bioplastics 

more soluble in water, susceptible to enzymatic attack by microorganisms, and opaque. 

Through of the contact angle test, the bioplastic with starch and plasticizer showed lower 

affinity to water compared to the one including xylan, which among them the proportion of 25 

% (w/w of the polysaccharides) was the least hydrophilic. Bioplastic with 5 % (w/w of 

polysaccharides) xylan showed better crystallinity followed by starch. As for its mechanical 

strength, the bioplastic with 10 % xylan (w/w of polysaccharides) owned the highest tensile 

stress (2.56 MPa), but the presence of xylan considerably reduced its elongation at break 

(ranging from 16 to 37 %) in relation to the starch bioplastic (208.8 %). In general, after 30 

days of burial, the bioplastics had practically no fragments visible to the naked eye, and the 

rate of CO2 production together with the increase in cell count after the biodegradation period 

demonstrate that there was an effective catabolic action of the microorganisms in the 

bioplastics. Thus, the 5 % xylan bioplastic showed the best physicochemical results when 

obtaining a biodegradable material. 

 

Keywords: Bioplastic, xylan, starch, biomass, biodegradable. 

 

 

 

 

 

 

 

 

 

 

 

 

 

 



 

 

RESUMO 

 

O plástico serviu muito bem à sociedade durante décadas, podendo se apresentar como um 

material leve e moldável, mecanicamente e termicamente resistente, inerte a produtos 

químicos, isolante elétrico, além de possuir características ópticas interessantes como sua 

transparência. Tais características, associada ao baixo preço da sua matéria prima, um resíduo 

do processamento de petróleo até então pouco utilizado, fizeram com que ele se tornasse 

crucial para a indústria e o mercado, o que consolidou sua presença na sociedade em 

diferentes países. Contudo, por ser um material resistente, permanecendo séculos sem se 

decompor totalmente, e por ter uma larga produção a fim de atender as necessidades de uma 

sociedade consumista, ele passou a se acumular no meio ambiente ao longo dos anos desde 

sua criação. Sua abundancia no meio ambiente acarreta prejuízos à vida em diferentes biomas, 

especialmente os ambientes marinhos, além de possuir resíduos que podem ser tóxicos e 

chegar ao ser humano através da cadeia trófica. Uma possível solução são os materiais 

biodegradáveis e que utilizam matéria-prima renovável como proteínas, lipídeos e 

polissacarídeos, os chamados bioplásticos. Neste contexto, o presente estudo objetivou 

produzir bioplásticos baseado em amido com diferentes proporções de xilana extraída do 

bagaço de cana-de-açúcar e glicerol como plastificante.  Para isso, suas propriedades físico-

químicas (solubilidade, hidrofilicidade, umidade, barreira à água, opacidade, cristalinidade, e 

resistência mecânica) foram avaliadas.  A biodegradaçãodo bioplástico foi determinada por 

enterramento e liberação de CO2. Como resultado, foi observado que o aumento na proporção 

de xilana tornou os bioplásticos mais solúveis em água, susceptíveis ao ataque enzimático de 

microrganismo, e opacos. Por meio do ensaio de ângulo de contato, o bioplástico apenas com 

amido e plastificante apresentou menor afinidade à água, comparado aos compostos por 

xilana, que dentre eles a proporção de 25 % (m/m de polissacarídeos) foi a menos hidrofílica. 

O bioplástico com 5 % (m/m de polissacarídeos) de xilana apresentou melhor cristalinidade 

seguido pelo de amido. Quanto à sua resistência mecânica, o bioplastico com 10 % de xilana 

(m/m de polissacarídeos) foi o que possuiu maior tensão de tração (2,56 MPa), porém a 

presença de xilana reduziu consideravelmente sua elongação na quebra (variando de 16 a 37 

%) em relação ao bioplástico de amido (208,8 %). No geral, após 30 dias enterrados os 

bioplásticos já não apresentavam praticamente nenhum fragmento visível a olho nú, e a taxa 

de produção de CO2 juntamente com o aumento da contagem de células após o período de 

biodegração demonstram que houve uma efetiva ação catabólica dos microrganimos nos 

bioplasticos. Sendo assim, a concentração de 5 % de xilana foi a que mostrou melhores 

resultados físico-químicos ao obter um material biodegradável.  

 

Palavras-chave: Bioplastico, xilana, amido, biomassa, biodegradável. 
 

 

 

 

 

 

 

 

 

 

 

 



 

 

LIST OF FIGURES 

 

Figure 1 - Polymeric chain models representing a linear polymer and a branched one. .......... 16 

Figure 2 - Illustration of a semi-rigid three-dimensional network structure. ........................... 27 

Figure 3 - Types of copolymers. ............................................................................................... 28 

Figure 4 - Groups of hemicellulose. ......................................................................................... 34 

Figure 5 - Drug-carrying biomaterial applied in tissue regeneration. ...................................... 36 

Figure 6 - Presence of plasticizer in polymers. ........................................................................ 39 

Figure 7 - Interaction between chitosan and xylan. .................................................................. 42 

Figure 8 - Esterification reaction. ............................................................................................. 45 

Figure 9 - Etherification reaction.............................................................................................. 46 

Figure 10 - Oxidation reaction.................................................................................................. 46 

Figure 11 - Chains crosslinked. ................................................................................................ 47 

Figure 12 - Examples of bioplastics produced from starch and xylan in different proportions 

in relation to 5 % of polysaccharides (w/w of filmogenic solution). ....................................... 80 

Figure 13 - X-ray diffractograms of bioplastic of starch and xylan (according to Table 3). ... 81 

Figure 14 - Relative percentage of contact angle of bioplastics of starch and xylan in relation 

to starch bioplastics. ................................................................................................................. 84 

Figure 15 - Opacity as a function of the composition of xylan in bioplastics, showing the 

respective photo of each composition ...................................................................................... 85 

Figure 16 - FTIR of starch and xylan bioplastics (according to Table 3). ............................... 86 

Figure 17 - Scanning electron microscopy images of starch and xylan bioplastics with starch 

and xylan with different magnitudes. ....................................................................................... 89 

Figure 18 - CO2 production (mg) from the biodegradation of starch and xylan bioplastics. ... 91 

Figure 19 - Decomposition of the bioplastic with 10% of xylan (X10) over 26 days. ............. 92 

Figure 20 - Plate growth of soil microorganisms of the mixture of soils used in each bioplastic 

composition .............................................................................................................................. 93 

 

 

 

 

 

 

 

 



 

 

LIST OF TABLES 

 

Table 1 - Evaluation of different proportions of glycerol (G) in 5 % starch (w/w of solution) 

bioplastics ................................................................................................................................. 73 

Table 2 - Evaluation of starch gelatinization conditions in bioplastics with 20 % glycerol (w/w 

of starch) ................................................................................................................................... 73 

Table 3 - Bioplastic compositions in relation to 5 % (w/w, filmogenic solution) of 

polysaccharides with glycerol 20 % (w/w, polysaccharides). .................................................. 74 

Table 4 - Xylan extraction yield (%, w/w theoretically present in the bagasse) ...................... 78 

Table 5 - Lignin content and sugars in biomass and solubilized xylan (%, w/w) .................... 78 

Table 6 - Moisture and solubility results of bioplastics with 5 % starch (w/w of solution) and 

glycerol. .................................................................................................................................... 80 

Table 7 - Solubility and moisture of bioplastics of starch and xylan (according to Table 3)... 82 

Table 8 - Water vapor transmission (WPT) and water vapor permeability (WVP) of 

bioplastics of starch and xylan (according to Table 3). ............................................................ 83 

Table 9 - Tensile stress at maximum load and elongation at break in starch and xylan 

bioplastics (according to Table 3). ........................................................................................... 88 

Table 10 - Average number of colony-forming units per gram of soil (CFU.g-1) before and 

after biodegradation of bioplastics (composed of the mixture of soils used in each 

composition). ............................................................................................................................ 93 

 

LIST OF APPENDIX 

 

Appendix A - Representative figure of the setup of the experiment witch a bioplastic 25 % 

xylan (w/w polysaccharides) .................................................................................................. 102 

Appendix B - Representation of the biodegradation test of bioplastics with variations of xylan 

and only starch. ....................................................................................................................... 103 

Appendix C - Interference of gelatinization in xylan and starch bioplastic (left pair), and only 

starch (right pair). ................................................................................................................... 104 

Appendix D - Bioplastics made in different ways and conditions of gelatinization. ............. 105 

 

 

 

 

 

 

 



 

 

SUMMARY 

CHAPTER I: INTRODUCTION ............................................................................................. 11 

CHAPTER II: OBJECTIVES ................................................................................................... 13 

1. General objective ................................................................................................................ 13 

1.1. Specific objectives ............................................................................................................. 13 

CHAPTER III: HEMICELLULOSE APPLICATION FOR THE PRODUCTION OF 

BIOPLASTICS AND BIOMATERIALS ................................................................................ 14 

1. Introduction ........................................................................................................................ 15 

2. Plastic ................................................................................................................................... 16 

2.1. Origin of plastics and their importance for society ........................................................... 16 

2.2. Negative impacts caused by an irregular plastics disposal ................................................ 18 

3. Bioplastics ............................................................................................................................ 25 

3.1. Origin of bioplastics .......................................................................................................... 25 

3.2. Bioplastic composition and structure ................................................................................ 27 

3.3. The bioplastics market ....................................................................................................... 35 

3.4. Emerging technologies for bioplastics produced with hemicellulose ............................... 35 

3.4.1. Importance of plasticizer for hemicellulose films .......................................................... 38 

3.4.2. Development of biomolecule blends ............................................................................... 40 

3.4.3. Crosslinking agents in polymeric chains ........................................................................ 47 

3.4.4. Functional biopackages .................................................................................................. 48 

3.4.5. Other applications of hemicellulose as a biomaterial .................................................... 51 

4. Concluding remarks ........................................................................................................... 53 

REFERENCES ......................................................................................................................... 55 

CHAPTER IV: DEVELOPMENT OF BIOPLASTICS BASED ON STARCH AND XYLAN: 

A MECHANICAL PHYSICOCHEMICAL AND BIODEGRADABILITY STUDY ............ 69 

1. Introduction ........................................................................................................................ 70 

2. Material and methods ........................................................................................................ 71 

2.1. Raw material ...................................................................................................................... 72 

2.2. Xylan solubilization ........................................................................................................... 72 

2.3. Characterization of biomass and extracted xylan .............................................................. 72 

2.4. Gelatinization parameters and plasticizer content ............................................................. 73 

2.5. Bioplastic production ......................................................................................................... 73 



 

2.6. Bioplastics characterization ............................................................................................... 74 

2.6.1. Moisture and solubility tests ........................................................................................... 74 

2.6.2. Opacity................ ........................................................................................................... 74 

2.6.3. Hydrophilicity ................................................................................................................. 75 

2.6.4. Water vapor transmission (WVT) and water vapor permeability (WVP) ...................... 75 

2.6.5. Mechanical test ............................................................................................................... 75 

2.6.6. Fourier-transform infrared spectroscopy (FTIR)........................................................... 76 

2.6.7. Scanning electron microscopy (SEM) ............................................................................ 76 

2.6.8. X-ray diffraction (XRD) .................................................................................................. 76 

2.7. Biodegradation .................................................................................................................. 76 

2.8. Statistical analysis ............................................................................................................. 77 

3. Results and discussion ........................................................................................................ 77 

3.1. Biomass composition, xylan solubilization, and characterization ..................................... 77 

3.2. Gelatinization parameters and plasticizer content ............................................................. 79 

3.3. Bioplastics characterization ............................................................................................... 81 

3.3.1. X-ray diffraction (XRD) .................................................................................................. 81 

3.3.2. Moisture and solubility tests ........................................................................................... 82 

3.3.3. Water vapor transmission (WVT) and water vapor permeability (WVP) ...................... 82 

3.3.4. Hydrophilicity ................................................................................................................. 83 

3.3.5. Opacity............................................................................................................................ 84 

3.3.6. Fourier-transform infrared spectroscopy (FTIR)........................................................... 85 

3.3.7. Mechanical test ............................................................................................................... 86 

3.3.8. Scanning electron microscopy (SEM) ............................................................................ 88 

3.4. Biodegradation .................................................................................................................. 89 

4. Conclusions ......................................................................................................................... 93 

REFERENCES ......................................................................................................................... 95 

CHAPTER V: CONCLUSION .............................................................................................. 101 

APPENDIX ............................................................................................................................ 102 

 

 

 

 

 

 

 

 

 



11 

CHAPTER I: INTRODUCTION 

The fossil-based plastic, that we conventionally use, assumed an important role in 

society in a way that it would be impossible to abruptly abolish its use, since it is present in 

our daily lives from the time we wake up to the time we go to sleep. Although plastic waste 

management policies can be carried out, the ideal level of recycling to contain the generation 

of waste, which tends to grow as the world population grows, is a difficult level to be reached. 

And the reasons for this are the demand for high investment in collection and sorting centers, 

the problem of the quality of recycled materials in relation to primary plastic and the most 

important factor, the cooperation of the population, because even if the first two items are 

resolved, will not be as effective if there is no awareness of the population. This last 

requirement is a more complex issue, because it is strictly related to the education and civility 

of each individual. 

Biodegradable polymers produced from renewable sources have been seen as a 

promising strategy to replace (partially) the intensive use of plastic derived from petroleum 

due to the growing environmental awareness in recent decades, as this synthetic polymer 

remains in the environment for centuries. The use of polysaccharides has been studied and 

arouses the interest of researchers as they are molecules capable of forming long chains. The 

polysaccharides, can form a cohesive network due to inter and intra-molecular bonds, and can 

also be consumed and degraded by microorganisms resulting in small molecules common to 

the environment. In addition to being biodegradable, bioplastics have the raw material from 

renewable sources, and potentially toxic substances are not used in their composition. By 

relying on several hydroxyl groups, polysaccharides also have the ability to mix with other 

components (plasticizers and additives) improving their functionality. 

Starch is generally used in bioplastic/film production because it is highly available and 

easy to extract, it has low cost, is biodegradable and biocompatible. It is still widely studied as 

edible bio-packaging because it has high energy value, in addition to forming films without 

odor, taste, color, toxicity, showing semipermiability to gases such as CO2, O2 and water 

vapor. However, native starch has a hydrophilic character and tends to form films with poor 

mechanical properties. Basically formed by two types of molecules, amylose (forms linear 

chains) and amylopectin (branched chains), both having a large amount of carboxyl groups 

capable of making hydrogen bonds.  



12 

In this context, xylan, a type of hemicellulose composed mostly by pentoses known as 

xylose, can present itself as a compatible sugar to starch, as it shares practically the same 

characteristics. Xylan can contribute to form bioplastic with greater mechanical resistance, in 

addition to theoretically preventing the retrogression process characteristic of starch, which 

consists of the loss of water molecules causing dryness. The interest in xylan also comes from 

the fact that it is a polysaccharide with potential for application in bioplastics that is still little 

used when compared to cellulose, even though it has considerable participation in the 

composition of plant biomass.  

The study of starch in the preparation of biodegradable bioplastic is already well 

documented. However, from our knowledge, there are not study in the literature dedicated to 

understanding the contribution and correlation of xylan in bioplastic. In this context, the 

purpose of this study was to combine starch and xylan (extracted from sugarcane bagasse) to 

produce bioplastics. The contribution of the xylan concentration was evaluated in properties 

such as biodegradability, crystallinity, mechanics, opacity, water affinity, and water barrier.  



13 

CHAPTER II: OBJECTIVES 

1. General objective

The main objective of this study was to evaluate the addition of xylan extracted from 

sugarcane bagasse in the composition of bioplastics in combination with starch.  

Physicochemical properties were determined and related to de biodegradation of the 

bioplastic. 

1.1. Specific objectives 

 Identify the better starch gelatinization conditions and glycerol concentration;

 Study the contribution of the xylan in the bioplastic compositions on the moisture,

solubility, opacity, mechanical, crystallinity, and hydrophilicity, defining the most

suitable proportion;

 Analyze the time needed for biodegradation of the bioplastic composed of xylan and

starch by burial and CO2 release.

. 



95 

 

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CHAPTER V: CONCLUSION 

 

Researchers from companies and governmental initiatives around the world are trying 

to develop a material that is similar to plastic but that can be completely degraded. In this 

sense, the use of biopolymers/biomolecules from plant biomass is a renewable and 

environmentally beneficial option in the production of bioplastics, as in the case of the xylan 

used in this study together with starch. In relation to the bioplastic production, the 

gelatinization process contributed to its integrity, as well as its flexibility, which was also 

favored when the glycerol content was 20 % (w/w of polysaccharides), resulting in films that 

were easier to handle and remove from the drying plate. The association of polysaccharides 

generated homogeneous and intact bioplastics, indicating a good association between them. 

Moreover, xylan tended to increase the opacity, solubility, and hydrophilicity of bioplastics 

reducing their crystallinity and elongation at break. When buried, the bioplastics were almost 

completely degraded within a month, and in those where xylan was more present, CO2 

production peaked in fewer days, indicating an even more accelerated decomposition. Its 

properties may not make xylan bioplastics ideal candidates for use in food packaging and 

bags, for example, but in applications that do not require high mechanical strength but rather a 

fast biodegradation rate, they are an efficient option, such as in agricultural applications where 

some products or even the cultivation itself needs a covering that preferably can be degraded 

over time without generating potentially toxic waste and plastic waste.